WO2002066504A2 - Protein-protein interactions in saccharomyces cerevisiae - Google Patents

Abstract

The present invention relates to proteins that interact with other proteins of Saccharomyces cerevisiae. More specifically, the present invention relates to complexes of polypeptides or polynucleotides encoding the polypeptides, fragments of the polypeptides, antibodies to the complexes, Selected Interacting Domains (SID®) which are identified due to the protein-protein interactions, methods for screening for agents which modulate the interaction of proteins and compositions that are capable of modulating the protein-protein interactions, such as, for example, drug in pharmaceutical composition.

Description

MORE PROTEIN-PROTEIN INTERACTIONS IN Saccharomyces cerevisiae

The present invention claims priority from U.S. provisional application No. 60/269,266 filed on February 16, 2002.

BACKGROUND Most biological processes involve specific protein-protein interactions. Protein-protein interactions enable two or more proteins to associate. A large number of non-covalent bonds form between the proteins when two protein surfaces are precisely matched. These bonds account for the specificity of recognition. Thus, protein-protein interactions are involved, for example, in the assembly of enzyme subunits, in antibody-antigen recognition, in the formation of biochemical complexes, in the correct folding of proteins, in the metabolism of proteins, in the transport of proteins, in the localization of proteins, in protein turnover, in first translation modifications, in the core structures of viruses and in signal transduction.

General methodologies to identify interacting proteins or to study these interactions have been developed. Among these methods are the two-hybrid system originally developed by Fields and co-workers and described, for example, in U.S. Patent Nos. 5,283,173, 5,468,614 and 5,667,973, which are hereby incorporated by reference.

The earliest and simplest two-hybrid system, which acted as basis for development of other versions, is an in vivo assay between two specifically constructed proteins. The first protein, known in the art as the "bait protein" is a chimeric protein which binds to a site on DNA upstream of a reporter gene by means of a DNA-binding domain or BD. Commonly, the binding domain is the DNA-binding domain from either Gal4 or native E. coli LexA and the sites placed upstream of the reporter are Gal4 binding sites or LexA operators, respectively.

The second protein is also a chimeric protein known as the "prey" in the art. This second chimeric protein carries an activation domain or AD. This activation domain is typically derived from Gal4, from VP16 or from B42. Besides the two hybrid systems, other improved systems have been developed to detected protein-protein interactions. For example, a two-hybrid plus one system was developed that allows the use of two proteins as bait to screen available cDNA libraries to detect a third partner. This method permits the detection between proteins that are part of a larger protein complex such as the RNA polymerase II holoenzyme and the TFIIH or TFIID complexes. Therefore, this method, in general, permits the detection of ternary complex formation as well as inhibitors preventing the interaction between the two previously defined fused proteins. Another advantage of the two-hybrid plus one system is that it allows or prevents the formation of the transcriptional activator since the third partner can be expressed from a conditional promoter such as the methionine-repressed Met25 promoter which is positively regulated in medium lacking methionine. The presence of the methionine-regulated promoter provides an excellent control to evaluate the activation or inhibition properties of the third partner due to its "on" and "off switch for the formation of the transcriptional activator. The three-hybrid method is described, for example in Tirade et al., The Journal of Biological Chemistry, 272, No. 37 pp. 22995-22999 (1997). incorporated herein by reference.

Besides the two and two-hybrid plus one systems, yet another variant is that described in Vidal et al, Proc. Natl. Sci. 93 pgs. 10315-10320 called the reverse two- and one-hybrid systems where a collection of molecules can be screened that inhibit a specific protein-protein or protein/DNA interactions, respectively.

A summary of the available methodologies for detecting protein-protein interactions is described in Vidal and Legrain, Nucleic Acids Research Vol. 27, No. 4 pgs.919-929 (1999) and Legrain and Selig, FEBS Letters 480 pgs. 32-36 (2000) which references are incorporated herein by reference.

However, the above conventionally used approaches and especially the commonly used two-hybrid methods have their drawbacks. For example, it is known in the art that, more often than not, false positives and false negatives exist in the screening method. In fact, a doctrine has been developed in this field for interpreting the results and in common practice an additional technique such as co-immunoprecipitation or gradient sedimentation of the putative interactors from the appropriate cell or tissue type are generally performed. The methods used for interpreting the results are described by Brent and Finley, Jr. in Ann. Rev. Genet, 31 pgs. 663-704 (1997). Thus, the data interpretation is very questionable using the conventional systems.

One method to overcome the difficulties encountered with the methods in the prior art is described in WO 99/42612, incorporated herein by reference. This method is similar to the two-hybrid system described in the prior art in that it also uses bait and prey polypeptides. However, the difference with this method is that a step of mating at least one first haploid recombinant yeast cell containing the prey polypeptide to be assayed with a second haploid recombinant yeast cell containing the bait polynucleotide is performed. Of course the person skilled in the art would appreciate that either the first recombinant yeast cell or the second recombinant yeast cell also contains at least one detectable reporter gene that is activated by a polypeptide including a transcriptional activation domain. The method described in WO 99/42612 permits the screening of more prey polynucleotides with a given bait polynucleotide in a single step than in the prior art systems due to the cell to cell mating strategy between haploid yeast cells. Furthermore, this method is more thorough and reproducible, as well as sensitive. Thus, the presence of false negatives and/or false positives is extremely minimal as compared to the conventional prior art methods.

Yeast are unicellular eukaryotic organisms; the yeast genus Saccharomyces comprises strains whose biochemistry and genetics are intensively studied in the laboratory; it also comprises strains frequently used in the industry, in particular in the food industry (bread, alcoholic drinks, etc.), and consequently produced in very large quantities.

Among the numerous applications involving Saccharomyces cerevisiae, the most frequent are:

Bread production. Production of ethanol or ethyl alcohol (C₂-H₅-OH) by fermentation methods.

Fermentation techniques for ethanol production developed during the early part of this century were supplemented by synthetic processes based on crude petroleum^", as oil was much cheaper and abundantly available. However, the fermentative production of ethanol has again picked up, using various kinds of renewable fermentable substrates, such as: (i) sugar (from sugar-cane, sugar beet, fruit) which may be converted to ethanol directly; (ii) starch (from grain, root crops) which is first hydrolysed to fermentable sugars by enzymes; and (iii) cellulose (from wood, agricultural wastes, etc.) which is converted to sugars. (Biotechnology: Economic and Social Aspects-Issues for Developing Countries, Eds. E. J. Da Silva, C. Ratiedge and A Sesson; Cambridge University Press, p.24, 1992). Ethanol production by fermentation is based mainly on yeast, and for large scale fuel production, these are generally of the genus Saccharomyces. (3) Additives

Yeast extract may provide product containing a large quantity of enzymes, coenzymes, ferments, group B vitamins, nucleotides, nucleosides, free amino acids and RNA acid. Particularly useful products are obtained from Saccharomyces cerevisiae strains having high resistance in an acid environment (e g. gastric juices) and towards antibiotics. These product characteristics make it particularly suitable and effective as a human and animal food additive, as a growth factor and intestinal bacterial flora regulator. Its action is both prophylactic and curative in many affections in the human and veterinary field deriving from enzymatic and bacterial imbalance of the intestine.

Production of proteins. The ease with which the genetics of Saccharomyces cerevisiae cells may be manipulated and the long industrial history of this species hence make it a host of choice for the production of foreign proteins using recombinant DNA techniques. Yeast cells have proven useful as hosts for production of heterologous gene products.

Yeast such as the bakers yeast Saccharomyces cerevisiae can be grown to high cell densities inexpensively in simple media, and helpful genetic techniques and molecular genetic methods are available. Accordingly, pharmaceutical preparations of human alpha-1-antitrypsin, and vaccines for hepatitis B virus have been produced in the cytoplasm of yeast cells and isolated by lysis of cells and purification of the desired protein (Valenzuela, P., et al., 1982, Nature 298: 347-350; Travis, J., et al., 1985, J. Biol. Chem. 260: 4384-4389). However, some proteins, such as prochymosin and prourokinase (also known as single-chain urinary plasminogen activator, or scu-PA) are produced much more efficiently by secretion from yeast cells, apparently because they are normally secreted from their native host cells and because proper folding of the polypeptide chain and disulfide bond formation occur only in the secretion pathway (Smith, Duncan, & Moir, 1985, Science 229: 1219-1224; Moir et al., 1988, Abstract 19 from The Ninth International Congress on Fibrinolysis, Amsterdam, The Netherlands).

This application needs improvement especially concerning the secretion of the protein of interest: The secreted yield of protein product is dependent upon both the gene to be expressed and the promoter and signal sequences chosen for its expression (Hitzeman, R. A., Leung, D. W., Perry, L. J., Kohr, W. J., Levine, H. L. and Goeddel, D. V. (1983) Science 219, 620-625; Bitter, G. A., Chen, K. K., Banks, A. R. and Lai, P.-H. (1984) Proc. Natl. Acad. Sci. USA 81 , 5330-5334; Brake, A. J., Merryweather, J. P., Coit, D. G., Heberlein, U. A., Masiarz, F. R., Mullenbach, G. T., Urdea, M. S., Valenzuela, P. and Barr, P. J. (1984) Proc. Natl. Acad. Sci. USA 81 , 4642-4646; Brake, A. J., Cousens, L. S., Urdea, M. S., Valenzuela, P. D. T. (1984) European Patent Application Publication No. 0 121 884). Although it is usually possible to obtain reasonably good production levels for a particular protein, often only a small fraction of the total amount produced can actually be found free in the medium. Most of the protein remains trapped inside the cell, often in the intracellular vacuole found in this species. In yeast, secretion can be regarded as a branched pathway with some secreted yeast proteins being "secreted" into the vacuole and others being directed across the plasma membrane to the periplasm and beyond (Sheckman, R. and Novick, P., in Strathern, J. N., Jones, E. W. and Broach, J. R. (eds.), Molecular Biology of the yeast Saccharomyces cerevisiae, Cold Springs Harbor Laboratory, Cold Springs Harbor, New York, 1981 , pp. 361-398). Apparently, some protein products of foreign genes are directed into the vacuolar branch of this pathway.

Yields of secreted heterologous proteins from yeast fermentations have been limited. Most non-yeast proteins are secreted quite inefficiently from yeast cells. For example, in all of the following cases, at least as much of the heteroloqous protein is found inside the cell as is found outside the cell in the culture broth or between the cell membrane and wall. This is true for calf prochymosin (Smith, Duncan, & Moir, 1985, Science 229: 1219-1223), human alpha-1- antitrypsin (Moir & Dumais, 1987, Gene 56: 209-217), human tissue plasminogen activator (Lemontt et al., 1985, DNA 4: 419-428), anchor-minus influenza hemagglutinin (Jabbar & Nayak, 1987, Mol. Cell. Biol. 7: 1476-1485), alpha interferon (Hitzeman et al., 1983, Science 219: 620-625), a consensus interferon (Zsebo et al., 1986, J. Biol. Chem. 261 : 5858-5865), murine lambda and mu immunoglobulin chains (Wood et al., 1985, Nature 314: 446-449), and human lysozyme (Jigami et al., 1986, Gene 43: 273-279). Clearly, methods are needed to increase the efficiency of secretion of these proteins and other non-yeast proteins from yeast cells. Such methods would provide therapeutic and industrially useful proteins more economically.

Model organism for medical study. S. cerevisiae is a research model organism for Candida infection study. It is also a model organism for human diseases, especially mechanisms involved in cancer (DNA repair, apoptose), neurodegenerative disease. The two last fields of application request a precise knowledge of yeast pathways, that is why there is a great need for construction of protein interaction map (PIM®) of Saccharomyces cerevisiae and to identify protein function in pathways of interest.

SUMMARY OF THE INVENTION The present invention relates to identifying protein-protein interactions for

Saccharomyces cerevisiae.

The present invention also relates to identifying protein-protein interactions of Saccharomyces cerevisiae for the development of more effective and better targeted therapeutic applications, for the development of yeast strain having a better secretion yield of protein of interest production (i. e., expression and secretion).

The present invention is also aimed at identifying complexes of polypeptides or polynucleotides encoding the polypeptides and fragments of the polypeptides of Saccharomyces cerevisiae.

Also, the present invention to identifying antibodies to these complexes of polypeptides or polynucleotides encoding the polypeptides and fragments of the polypeptides of Saccharomyces cerevisiae including polyclonal, as well as monoclonal antibodies that are used for detection.

The present invention also concerns the identification of selected interacting domains of the polypeptides, called SID® polypeptides. Furthermore, the present invention concerns the identification of selected interacting domains of the polynucleotides, called SID® polynucleotides.

The present invention is aimed at generating protein-protein interactions maps called PIM®s.

Also the present invention concerns a method for screening drugs for agents which modulate the interaction of proteins and pharmaceutical compositions that are capable of modulating the protein-protein interactions of Saccharomyces cerevisiae. The present invention also relates to administering the nucleic acids of the present invention via gene therapy.

Also, the present invention provides protein chips or protein microarrays.

In another embodiment the present invention provides a report in, for example paper, electronic and/or digital forms, concerning the protein-protein interactions, the modulating compounds and the like as well as a PIM®.

Moreover, the present invention relates to a protein complex of Saccharomyces cerevisiae.

The present invention also provides antibodies to the protein-protein complexes for Saccharomyces cerevisiae.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 is a schematic representation of the pB1 plasmid. Fig. 2 is a schematic representation of the pB5 plasmid. Fig. 3 is a schematic representation of the pB6 plasmid. Fig. 4 is a schematic representation of the pB13 plasmid. Fig. 5 is a schematic representation of the pB14 plasmid. Fig. 6 is a schematic representation of the pB20 plasmid. Fig. 7 is a schematic representation of the pP1 plasmid. Fig. 8 is a schematic representation of the pP2 plasmid. Fig. 9 is a schematic representation of the pP3 plasmid. Fig. 10 is a schematic representation of the pP6 plasmid. Fig. 1 1 is a schematic representation of the pP7 plasmid. Fig. 12 is a schematic representation of vectors expressing the T25 fragment. Fig. 13 is a schematic representation of vectors expressing the T18 fragment.

Fig. 14 is a schematic representation of various vectors of pCmAHLI , pT25 and pT18. Fig. 15 is a schematic representation identifying a SID®. In this figure the "Full-length prey protein" is the Open Reading Frame (ORF) or coding sequence (CDS) where the identified prey polypeptides are included. The Selected Interaction Domain (SID®) is determined by the commonly shared polypeptide domain of every selected prey fragment. Fig. 16 is an embodiment of a protein interaction map (PIM®).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

As used herein the terms "polynucleotides", "nucleic acids" and "oligonucleotides" are used interchangeably and include, but are not limited to RNA, DNA, RNA/DNA sequences of more than one nucleotide in either single chain or duplex form. The polynucleotide sequences of the present invention may be prepared from any known method including, but not limited to, any synthetic method, any recombinant method, any ex vivo generation method and the like, as well as combinations thereof.

The term "polypeptide" means herein a polymer of amino acids having no specific length. Thus, peptides, oligopeptides and proteins are included in the definition of "polypeptide" and these terms are used interchangeably throughout the specification, as well as in the claims. The term "polypeptide" does not exclude post-translational modifications such as polypeptides having covalent attachment of glycosyl groups, acetyl groups, phosphate groups, lipid groups and the like. Also encompassed by this definition of "polypeptide" are homologs thereof.

By the term "homologs" is meant structurally similar genes contained within a given species, orthologs are functionally equivalent genes from a given species or strain, as determined for example, in a standard complementation assay. Thus, a polypeptide of interest can be used not only as a model for identifying similar genes in given strains, but also to identify homologs and orthologs of the polypeptide of interest in other species. The orthologs, for example, can also be identified in a conventional complementation assay. In addition or alternatively, such orthologs can be expected to exist in bacteria (or other kind of cells) in the same branch of the phylogenic tree, as set forth, for example, at ftp://ftp.cme.msu.edu/pub/rdp/SSU-rKNA/SSU/Prok.phylo.

As used herein the term "prey polynucleotide" means a chimeric polynucleotide encoding a polypeptide comprising (i) a specific domain; and (ii) a polypeptide that is to be tested for interaction with a bait polypeptide. The specific domain is preferably a transcriptional activating domain.

As used herein, a "bait polynucleotide" is a chimeric polynucleotide encoding a chimeric polypeptide comprising (i) a complementary domain; and (ii) a polypeptide that is to be tested for interaction with at least one prey polypeptide. The complementary domain is preferably a DNA-binding domain that recognizes a binding site that is further detected and is contained in the host organism.

As used herein "complementary domain" is meant a functional constitution of the activity when bait and prey are interacting; for example, enzymatic activity.

As used herein "specific domain" is meant a functional interacting activation domain that may work through different mechanisms by interacting directly or indirectly through intermediary proteins with RNA polymerase II or Ill-associated proteins in the vicinity of the transcription start site.

As used herein the term "complementary" means that, for example, each base of a first polynucleotide is paired with the complementary base of a second polynucleotide whose orientation is reversed. The complementary bases are A and T (or A and U) or C and G.

The term "sequence identity" refers to the identity between two peptides or between two nucleic acids. Identity between sequences can be determined by comparing a position in each of the sequences which may be aligned for the purposes of comparison. When a position in the compared sequences is occupied by the same base or amino acid, then the sequences are identical at that position. A degree of sequence identity between nucleic acid sequences is a function of the number of identical nucleotides at positions shared by these sequences. A degree of identity between amino acid sequences is a function of the number of identical amino acid sequences that are shared between these sequences. Since two polypeptides may each (i) comprise a sequence (i.e., a portion of a complete polynucleotide sequence) that is similar between two polynucleotides, and (ii) may further comprise a sequence that is divergent between two polynucleotides, sequence identity comparisons between two or more polynucleotides over a "comparison window" refers to the conceptual segment of at least 20 contiguous nucleotide positions wherein a polynucleotide sequence may be compared to a reference nucleotide sequence of at least 20 contiguous nucleotides and wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences.

To determine the percent identity of two amino acids sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison. For example, gaps can be introduced in the sequence of a first amino acid sequence or a first nucleic acid sequence for optimal alignment with the second amino acid sequence or second nucleic acid sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position.

The percent identity between the two sequences is a function of the number of identical positions shared by the sequences. Hence % identity = number of identical positions / total number of overlapping positions X 100.

In this comparison the sequences can be the same length or may be different in length. Optimal alignment of sequences for determining a comparison window may be conducted by the local homology algorithm of Smith and Waterman (J. Theor. Biol., 91 (2) pgs. 370-380 (1981 ), by the homology alignment algorithm of Needleman and Wunsch, J. Miol. Biol., 48(3) pgs. 443-453 (1972), by the search for similarity via the method of Pearson and Lipman, PNAS, USA, 85(5) pgs. 2444-2448 (1988) , by computerized implementations of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetic Computer Group, 575, Science Drive, Madison, Wisconsin) or by inspection. The best alignment (i.e., resulting in the highest percentage of identity over the comparison window) generated by the various methods is selected. The term "sequence identity" means that two polynucleotide sequences are identical (i.e., on a nucleotide by nucleotide basis) over the window of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over the window of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size) and multiplying the result by 100 to yield the percentage of sequence identity. The same process can be applied to polypeptide sequences. The percentage of sequence identity of a nucleic acid sequence or an amino acid sequence can also be calculated using BLAST software (Version 2.06 of September 1998) with the default or user defined parameter.

The term "sequence similarity" means that amino acids can be modified while retaining the same function. It is known that amino acids are classified according to the nature of their side groups and some amino acids such as the basic amino acids can be interchanged for one another while their basic function is maintained.

The term "isolated" as used herein means that a biological material such as a nucleic acid or protein has been removed from its original environment in which it is naturally present. For example, a polynucleotide present in a plant, mammal or animal is present in its natural state and is not considered to be isolated. The same polynucleotide separated from the adjacent nucleic acid sequences in which it is naturally inserted in the genome of the plant or animal is considered as being "isolated."

The term "isolated" is not meant to exclude artificial or synthetic mixtures with other compounds, or the presence of impurities which do not interfere with the biological activity and which may be present, for example, due to incomplete purification, addition of stabilizers or mixtures with pharmaceutically acceptable excipients and the like.

"Isolated polypeptide" or "isolated protein" as used herein means a polypeptide or protein which is substantially free of those compounds that are normally associated with the polypeptide or protein in a naturally state such as other proteins or polypeptides, nucleic acids, carbohydrates, lipids and the like. The term "purified" as used herein means at least one order of magnitude of purification is achieved, preferably two or three orders of magnitude, most preferably four or five orders of magnitude of purification of the starting material or of the natural material. Thus, the term "purified" as utilized herein does not mean that the material is 100% purified and thus excludes any other material. The term "variants" when referring to, for example, polynucleotides encoding a polypeptide variant of a given reference polypeptide are polynucleotides that differ from the reference polypeptide but generally maintain their functional characteristics of the reference polypeptide. A variant of a polynucleotide may be a naturally occurring allelic variant or it may be a variant that is known naturally not to occur. Such non-naturally occurring variants of the reference polynucleotide can be made by, for example, mutagenesis techniques, including those mutagenesis techniques that are applied to polynucleotides, cells or organisms. Generally, differences are limited so that the nucleotide sequences of the reference and variant are closely similar overall and, in many regions identical.

Variants of polynucleotides according to the present invention include, but are not limited to, nucleotide sequences which are at least 95% identical after alignment to the reference polynucleotide encoding the reference polypeptide. These variants can also have 96%, 97%, 98% and 99.999% sequence identity to the reference polynucleotide.

Nucleotide changes present in a variant polynucleotide may be silent, which means that these changes do not alter the amino acid sequences encoded by the reference polynucleotide.

Substitutions, additions and/or deletions can involve one or more nucleic acids. Alterations can produce conservative or non-conservative amino acid substitutions, deletions and/or additions.

Variants of a prey or a SID® polypeptide encoded by a variant polynucleotide can possess a higher affinity of binding and/or a higher specificity of binding to its protein or polypeptide counterpart, against which it has been initially selected. In another context, variants can also loose their ability to bind to their protein or polypeptide counterpart.

By "fragment of a polynucleotide" or fragment of a "SID® polynucleotide" is meant that fragments of these sequences have at least 12 consecutive nucleotides, or between 12 and 500 consecutive nucleotides or between 12 and 1 ,500 consecutive nucleotides or between 12 and 3,000 consecutive nucleotides. By "fragment of a polypeptide" or fragment of a "SID® polypeptide" is meant that fragments of these sequences have at least 4 consecutive amino acids, between 4 and 160 consecutive amino acids or between 4 and 500 consecutive amino acids or between 4 and 1 ,000 consecutive amino acids.

By "anabolic pathway" is meant a reaction or series of reactions in a metabolic pathway that synthesize complex molecules from simpler ones, usually requiring the input of energy. An anabolic pathway is the opposite of a catabolic pathway.

As used herein, a "catabolic pathway" is a series of reactions in a metabolic pathway that break down complex compounds into simpler ones, usually releasing energy in the process. A catabolic pathway is the opposite of an anabolic pathway. As used herein, "drug metabolism" is meant the study of how drugs are processed and broken down by the body. Drug metabolism can involve the study of enzymes that break down drugs, the study of how different drugs interact within the body and how diet and other ingested compounds affect the way the body processes drugs.

As used herein, "metabolism" means the sum of all of the enzyme-catalyzed reactions in living cells that transform organic molecules. By "secondary metabolism" is meant pathways producing specialized metabolic products that are not found in every cell.

As used herein, "SID®" means a Selected Interacting Domain and is identified as follows: for each bait polypeptide screened, selected prey polypeptides are compared. Overlapping fragments in the same ORF or CDS define the selected interacting domain. As used herein the term "PIM®" means a protein-protein interaction map. This map is obtained from data acquired from a number of separate screens using different bait polypeptides and is designed to map out all of the interactions between the polypeptides.

The term "affinity of binding", as used herein, can be defined as the affinity constant Ka when a given SID® polypeptide of the present invention which binds to a polypeptide and is the following mathematical relationship:

(a) [SID®/polypeptide complex] Ka =

(b) [free SID®] [free polypeptide] wherein [free SID®], [free polypeptide] and [SID®/polypeptide complex] consist of the concentrations at equilibrium respectively of the free SID® polypeptide, of the free polypeptide onto which the SID® polypeptide binds and of the complex formed between SID® polypeptide and the polypeptide onto which said SID® polypeptide specifically binds.

The affinity of a SID® polypeptide of the present invention or a variant thereof for its polypeptide counterpart can be assessed, for example, on a Biacore™ apparatus marketed by Amersham Pharmacia Biotech Company such as described by Szabo et al Curr Opin Struct S/^'ol

5 pgs. 699-705 (1995) and by Edwards and Leartherbarrow, Anal. Biochem 246 pgs. 1-6

(1997).

As used herein the phrase "at least the same affinity" with respect to the binding affinity between a SID® polypeptide of the present invention to another polypeptide means that the Ka is identical or can be at least two-fold, at least three-fold or at least five fold greater than the Ka value of reference.

As used herein, the term "modulating compound" means a compound that inhibits or stimulates or can act on another protein which can inhibit or stimulate the protein-protein interaction of a complex of two polypeptides or the protein-protein interaction of two polypeptides. More specifically, the present invention comprises complexes of polypeptides or polynucleotides encoding the polypeptides composed of a bait polypeptide, or a bait polynucleotide encoding a bait polypeptide and a prey polypeptide or a prey polynucleotide encoding a prey polypeptide. The prey polypeptide or prey polynucleotide encoding the prey polypeptide is capable of interacting with a bait polypeptide of interest in various hybrid systems.

As described in the Background of the present invention there are various methods known in the art to identify prey polypeptides that interact with bait polypeptides of interest. These methods, include, but are not limited to, generic two-hybrid systems as described by Fields et al in Nature, 340:245-246 (1989) and more specifically in U.S. Patent Nos. 5,283,173, 5,468,614 and 5,667,973, which are hereby incorporated by reference; the reverse two-hybrid system described by Vidal et al, supra; the two plus one hybrid method described, for example, in Tirode et al, supra; the yeast forward and reverse 'n'-hybrid systems as described in Vidal and Legrain, supra; the method described in WO 99/42612; those methods described in Legrain et al FEBS Letters 480 pgs. 32-36 (2000) and the like.

The present invention is not limited to the type of method utilized to detect protein- protein interactions and therefore any method known in the art and variants thereof can be used. It is however better to use the method described in WO 99/42612 or WO 00/66722, both references incorporated herein by reference due to the methods' sensitivity, reproducibility and reliability.

Protein-protein interactions can also be detected using complementation assays such as those described by Pelletier et al. at http://www.abrf.org/JBT/Articles/JBT0012/ibtOO 12.html. WO 00/07038 and WO98/34120.

Although the above methods are described for applications in the yeast system, the present invention is not limited to detecting protein-protein interactions using yeast, but also includes similar methods that can be used in detecting protein-protein interactions in, for example, mammalian systems as described, for example in Takacs et al., Proc. Natl. Acad. Sci., USA, 90 (21):10375-79 (1993) and Vasavada et al., Proc. Natl. Acad. Sci., USA, 88 (23):10686- 90 (1991 ), as well as a bacterial two-hybrid system as described in Karimova et al (1998), W099/28746, WO 00/66722 and Legrain et al FEBS Letters, 480 pgs. 32-36 (2000).

The above-described methods are limited to the use of yeast, mammalian cells and

Escherichia coli cells, the present invention is not limited in this manner. Consequently, mammalian and typically human cells, as well as bacterial, yeast, fungus, insect, nematode and plant cells are encompassed by the present invention and may be transfected by the nucleic acid or recombinant vector as defined herein.

Examples of suitable cells include, but are not limited to, VERO cells, HELA cells such as ATCC No. CCL2, CHO cell lines such as ATCC No. CCL61 , COS cells such as COS-7 cells and ATCC No. CRL 1650 cells, W138, BHK, HepG2, 3T3 such as ATCC No. CRL6361 , A549, PC12, K562 cells, 293 cells, Sf9 cells such as ATCC No. CRL1711 and Cv1 cells such as ATCC No. CCL70.

Other suitable cells that can be used in the present invention include, but are not limited to, prokaryotic host cells strains such as Escherichia coli, (e.g., strain DH5-α), Bacillus subtilis, Salmonella typhimurium, or strains of the genera of Pseudomonas, Streptomyces and Staphylococcus.

Further suitable cells that can be used in the present invention include yeast cells such as those of Saccharomyces such as Saccharomyces cerevisiae. The bait polynucleotide, as well as the prey polynucleotide can be prepared according to the methods known in the art such as those described above in the publications and patents reciting the known method per se.

The bait polynucleotide of the present invention is obtained from Saccharomyces cerevisiae genomic DNA, usually Open Reading Frame. The prey polynucleotide is obtained from Saccharomyces cerevisiae genomic DNA, variants of genomic DNA, and fragments from the genome or transcriptome of Saccharomyces cerevisiae ranging from about 200 nucleic acids to about 3000 nucleic acids. The prey polynucleotide is then selected, sequenced and identified.

A genomic DNA prey library is prepared from the Saccharomyces cerevisiae and constructed in the specially designed prey vector pP2 as shown in Figure 8 after ligation of suitable linkers such that every genomic DNA fragment insert is fused to a nucleotide sequence in the vector that encodes the transcription activation domain of a reporter gene. Any transcription activation domain can be used in the present invention. Examples include, but are not limited to, Gal4,YP16, B42, His and the like. Toxic reporter genes, such as CAT^R, CYH2, CYH1 , URA3, bacterial and fungi toxins and the like can be used in reverse two-hybrid systems.

The polypeptides encoded by the nucleotide inserts of the genomic DNA fragment prey library thus prepared are termed "prey polypeptides" in the context of the presently described selection method of the prey polynucleotides.

The bait polynucleotide can be inserted in bait plasmid pB6 as illustrated in Figure 3. The bait polynucleotide insert is fused to a polynucleotide encoding the binding domain of, for example, the Gal4 DNA binding domain and the shuttle expression vector is used to transform cells.

As stated above, any cells can be utilized in transforming the bait and prey polynucleotides of the present invention including mammalian cells, bacterial cells, yeast cells, insect cells and the like.

In an embodiment, the present invention identifies protein-protein interactions in yeast. In using known methods a prey positive clone is identified containing a vector which comprises a nucleic acid insert encoding a prey polypeptide which binds to a bait polypeptide of interest. The method in which protein-protein interactions are identified comprises the following steps:

(i) mating at least one first haploid recombinant yeast cell clone from a recombinant yeast cell clone library that has been transformed with a plasmid containing the prey polynucleotide to be assayed with a second haploid recombinant yeast cell clone transformed with a plasmid containing a bait polynucleotide encoding for the bait polypeptide;

(ii) cultivating diploid cell clones obtained in step i) on a selective medium; and

(iii) selecting recombinant cell clones which grow on the selective medium.

This method may further comprise the step of: characterizing the prey polynucleotide contained in each recombinant cell clone which is selected in step iii).

In yet another embodiment of the present invention, in lieu of yeast, Escherichia coli is used in a bacterial two-hybrid system, which encompasses a similar principle to that described above for yeast, but does not involve mating for characterizing the prey polynucleotide. In yet another embodiment of the present invention, mammalian cells and a method similar to that described above for yeast for characterizing the prey polynucleotide are used.

By performing the yeast, bacterial or mammalian two-hybrid system it is possible to identify for one particular bait an interacting prey polypeptide. The prey polynucleotide that has been selected by testing the library of preys in a screen using the two-hybrid, two plus one hybrid methods and the like, encodes the polypeptide interacting with the protein of interest.

The present invention is also directed, in a general aspect, to a complex of polypeptides, polynucleotides encoding the polypeptides composed of a bait polypeptide or bait polynucleotide encoding the bait polypeptide and a prey polypeptide or prey polynucleotide encoding the prey polypeptide capable of interacting with the bait polypeptide of interest. These complexes are identified in Table I.

In another aspect, the present invention relates to a complex of polynucleotides consisting of a first polynucleotide, or a fragment thereof, encoding a prey polypeptide that interacts with a bait polypeptide and a second polynucleotide or a fragment thereof. This fragment has at least 12 consecutive nucleotides, but can have between 12 and 500 consecutive nucleotides, or between 12 and 1 ,500 consecutive nucleotides or between 12 and 3,000 consecutive nucleotides.

In yet another embodiment, the present invention relates to an isolated complex of at least two polypeptides encoded by two polynucleotides wherein said two polypeptides are associated in the complex by affinity binding and are depicted in Table I. In yet another embodiment, the present invention relates to an isolated complex comprising at least a polypeptide as described in column 1 of Table I and a polypeptide as described in column 2 of Table I. The present invention is not limited to these polypeptide complexes alone but also includes the isolated complex of the two polypeptides in which fragments and/or homologous polypeptides exhibit at least 95% sequence identity, as well as from 96% sequence identity to 99.999% sequence identity.

Also encompassed in another embodiment of the present invention is an isolated complex comprising a polypeptide, or a nucleotide coding for this polypeptide (column 1 , Table II) and a SID® polypeptide (column 3, table II), or a polynucleotide coding for this SID® polypeptide (column 2 Table II) interacting with the polypeptide.

Besides the isolated complexes described above, polynucleotide coding for a Selected Interacting Domain (SID®) polypeptide or a variant thereof or any of the polynucleotide set forth in Table II can be inserted into an expression vector which contains the necessary elements for the transcription and translation of the inserted protein-coding sequence. Such transcription elements include a regulatory region and a promoter. Thus, the polynucleotide which may encode a marker compound of the present invention is operably linked to a promoter in the expression vector. The expression vector may also include a replication origin. A wide variety of host/expression vector combinations are employed in expressing the nucleic acids of the present invention. Useful expression vectors that can be used include, for example, segments of chromosomal, non-chromosomal and synthetic DNA sequences. Suitable vectors include, but are not limited to, derivatives of SV40 and pcDNA and known bacterial plasmids such as col El, pCR1 , pBR322, pMal-C2, pET, pGEX as described by Smith et al [need cite 1988], pMB9 and derivatives thereof, plasmids such as RP4, phage DNAs such as the numerous derivatives of phage I such as NM989, as well as other phage DNA such as M13 and filamentous single stranded phage DNA; yeast plasmids such as the 2 micron plasmid or derivatives of the 2m plasmid, as well as centomeric and integrative yeast shuttle vectors; vectors useful in eukaryotic cells such as vectors useful in insect or mammalian cells; vectors derived from combinations of plasmids and phage DNAs, such as plasmids that have been modified to employ phage DNA or the expression control sequences; and the like.

For example in a baculovirus expression system, both non-fusion transfer vectors, such as, but not limited to pVL941 (SamHI cloning site Summers, pVL1393 (Sa HI, S al, Xba\, EcoRI, Not\, Xma\\\, Bglll and Pst\ cloning sites; Invitrogen) pVL1392 (Sglll, Psfl, Not\, Xma\\\, EcoRI, Xbal\, Smal and SamHI cloning site; Summers and Invitrogen) and pBlueSaclll (SamHI, BglH, Pst\, Λ/col and Hind\\\ cloning site, with blue/white recombinant screening, Invitrogen), and fusion transfer vectors such as, but not limited to, pAc700(SamHI and Kpn\ cloning sites, in which the SamHI recognition site begins with the initiation codon; Summers), pAc701 and pAc70-2 (same as pAc700, with different reading frames), pAc360 (SamHI cloning site 36 base pairs downstream of a polyhedrin initiation codon; Invitrogen (195)) and pBlueBacHisA, B, C ( three different reading frames with SamHI, BglU, Pst\, Λ/col and Hind\\\ cloning site, an N- terminal peptide for ProBond purification and blue/white recombinant screening of plaques; Invitrogen (220) can be used.

Mammalian expression vectors contemplated for use in the invention include vectors with inducible promoters, such as the dihydrofolate reductase promoters, any expression vector with a DHFR expression cassette or a DHFR/methotrexate co-amplification vector such as pED (Psfl, Sa/I, Sbal, Smal and EcoRI cloning sites, with the vector expressing both the cloned gene and DHFR; Kaufman, 1991 ). Alternatively a glutamine synthetase/methionine sulfoximine co- amplification vector, such as pEE14 (H/noflll, Xball, Smal, Sbal, EcoRI and Bc/I cloning sites in which the vector expresses glutamine synthetase and the cloned gene; Celltech). A vector that directs episomal expression under the control of the Epstein Barr Virus (EBV) or nuclear antigen (EBNA) can be used such as pREP4 (SamHI, Sfil, Xhol, Notl, Nhel, Hindlll, Nhel, Pvu l and Kpnl cloning sites, constitutive RSV-LTR promoter, hygromycin selectable marker; Invitrogen) pCEP4 (SamHI, Sfil, Xhol, Notl, Nhel, Hindlll, Nhel, Pvull and Kpnl cloning sites, constitutive hCMV immediate early gene promoter, hygromycin selectable marker; Invitrogen), pMEP4 (Kpnl, Pvul, Nhel, Hindlll, Notl, Xhol, Sfil, SamHI cloning sites, inducible methallothionein lla gene promoter, hygromycin selectable marker, Invitrogen), pREPδ (SamHI, Xnol, Notl, Hindlll, Nhel and Kpnl cloning sites, RSV-LTR promoter, histidinol selectable marker; Invitrogen), pREP9 (Kpnl, Nhel, Hindlll, Notl, Xhol, Sfil, SamHI cloning sites, RSV-LTR promoter, G418 selectable marker; Invitrogen), and pEBVHis (RSV-LTR promoter, hygromycin selectable marker, N-terminal peptide purifiable via ProBond resin and cleaved by enterokinase; Invitrogen).

Selectable mammalian expression vectors for use in the invention include, but are not limited to, pRc/CMV (Hindlll, BstXl, Notl, Sbal and Apal cloning sites, G418 selection, Invitrogen), pRc/RSV (Hindll, Spel, BstXl, Notl, Xbal cloning sites, G418 selection, Invitrogen) and the like. Vaccinia virus mammalian expression vectors (see, for example Kaufman 1991 that can be used in the present invention include, but are not limited to, pSC11 (Smal cloning site, TK- and β-gal selection), pMJ601 (Sail, Smal, Afll, Naή, SspMII, SamHI, Apal, Nhel, Sacll, Kpnl and Hindlll cloning sites; TK- and β-gal selection), pTKgptFIS (EcoRI, Psfl, Salll, Accl, Hindll, Sbal, SamHI and Hpa cloning sites, TK or XPRT selection) and the like. Yeast expression systems that can also be used in the present include, but are not limited to, the non-fusion pYES2 vector (Xbal, Spnl, Snol, Notl, GstXl, EcoRI, BstXl, SamHI, Sad, Kpnl and H/noflll cloning sites, Invitrogen), the fusion pYESHisA, B, C (Xball, Sphl, Sho , Notl, BstXl, EcoRI, SamHI, Sacl, Kpnl and Hindlll cloning sites, N-terminal peptide purified with ProBond resin and cleaved with enterokinase; Invitrogen), pRS vectors and the like. Consequently, mammalian and typically human cells, as well as bacterial, yeast, fungi, insect, nematode and plant cells an used in the present invention and may be transfected by the nucleic acid or recombinant vector as defined herein. Examples of suitable cells include, but are not limited to, VERO cells, HELA cells such as ATCC No. CCL2, CHO cell lines such as ATCC No. CCL61 , COS cells such as COS-7 cells and ATCC No. CRL 1650 cells, W138, BHK, HepG2, 3T3 such as ATCC No. CRL6361 , A549,

PC12, K562 cells, 293 cells, Sf9 cells such as ATCC No. CRL1711 and Cv1 cells such as ATCC No. CCL70.

Other suitable cells that can be used in the present invention include, but are not limited to, prokaryotic host cells strains such as Escherichia coli, (e.g., strain DH5-α), Bacillus subtilis, Salmonella typhimurium, or strains of the genera of Pseudomonas, Streptomyces and Staphylococcus. Further suitable cells that can be used in the present invention include yeast cells such as those of Saccharomyces such as Saccharomyces cerevisiae.

Besides the specific isolated complexes, as described above, the present invention relates to and also encompasses SID® polynucleotides. As explained above, for each bait polypeptide, several prey polypeptides may be identified by comparing and selecting the intersection of every isolated fragment that are included in the same polypeptide. Thus the SID® polynucleotides of the present invention are represented by the shared nucleic acid sequences of uneven SEQ ID from n°1 to 863 (column 2, Table II) encoding the SID® polypeptides of even SEQ ID from n°2 to 864 (column 3 of Table II).

The present invention is not limited to the SID® sequences as described in the above paragraph, but also includes fragments of these sequences having at least 12 consecutive nucleic acids, or between 12 and 500 consecutive nucleic acids, or between 12 and 1 ,500 consecutive nucleic acids, or between 12 and 3,000 consecutive nucleic acids, as well as variants thereof. The fragments or variants of the SID® sequences possess at least the same affinity of binding to its protein or polypeptide counterpart, against which it has been initially selected. Moreover this variant and/or fragments of the SID® sequences alternatively can have between 95% and 99.999% sequence identity to its protein or polypeptide counterpart.

According to the present invention the variants can be created by known mutagenesis techniques either in vitro or in vivo. Such a variant can be created such that it has altered binding characteristics with respect to the target protein and more specifically that the variant binds the target sequence with either higher or lower affinity.

Polynucleotides that are complementary to the above sequences which include the polynucleotides of the SID®'s, their fragments, variants and those that have specific sequence identity are also included in the present invention.

The polynucleotide encoding the SID® polypeptide, fragment or variant thereof can also be inserted into recombinant vectors which are described in detail above. The present invention also relates to a composition comprising the above-mentioned recombinant vectors containing the SID® polynucleotides in Table II, fragments or variants thereof, as well as recombinant host cells transformed by the vectors. The recombinant host cells that can be used in the present invention were discussed in greater detail above. The compositions comprising the recombinant vectors can contain physiological acceptable carriers such as diluents, adjuvants, excipients and any vehicle in which this composition can be delivered therapeutically and can include, but is are not limited to sterile liquids such as water and oils.

In yet another embodiment, the present invention relates to a method of selecting modulating compounds, as well as the modulating molecules or compounds themselves which may be used in a pharmaceutical composition. These modulating compounds may act as a cofactor, as an inhibitor, as antibodies, as tags, as a competitive inhibitor, as an activator or alternatively have agonistic or antagonistic activity on the protein-protein interactions.

The activity of the modulating compound does not necessarily, for example, have to be 100% activation or inhibition. Indeed, even partial activation or inhibition can be achieved that is of pharmaceutical interest.

The modulating compound can be selected according to a method which comprises:

(i) cultivating a recombinant host cell with a modulating compound on a selective medium and a reporter gene the expression of which is toxic for said recombinant host cell wherein said recombinant host cell is transformed with two vectors: wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide having a DNA binding domain; wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide having a transcriptional activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact;

(ii) selecting said modulating compound which inhibits or permits the growth of said recombinant host cell.

Thus, the present invention relates to a modulating compound that inhibits the protein- protein interactions of a complex of two polypeptides as described in Table I. The present invention also relates to a modulating compound that activates the protein-protein interactions of a complex of two polypeptides as described in Table I.

In yet another embodiment, the present invention relates to a method of selecting a modulating compound, which modulating compound inhibits the interactions of two polypeptides as described in Table I. This method comprises: cultivating a recombinant host cell with a modulating compound on a selective medium and a reporter gene the expression of which is toxic for said recombinant host cell wherein said recombinant host cell is transformed with two vectors: wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide having a first domain of an enzyme; wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide having an enzymatic transcriptional activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact; selecting said modulating compound which inhibits or permits the growth of said recombinant host cell.

In the two methods described above any toxic reporter gene can be utilized including those reporter genes that can be used for negative selection including the URA3 gene, the CYH1 gene, the CYH2 gene and the like.

In yet another embodiment, the present invention provides a kit for screening a modulating compound. This kit comprises a recombinant host cell which comprises a reporter gene the expression of which is toxic for the recombinant host cell. The host cell is transformed with two vectors. The first vector comprises a polynucleotide encoding a first hybrid polypeptide having a DNA binding domain; and the second vector comprises a polynucleotide encoding a second hybrid polypeptide having a transcriptional activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact.

In yet another embodiment a kit is provided for screening a modulating compound by providing a recombinant host cell, as described in the paragraph above, but instead of a DNA binding domain, the first vector encodes a first hybrid polypeptide containing a first domain of a protein. The second vector encodes a second polypeptide containing a second part of a complementary domain of a protein that activates the toxic reporter gene when the first and second hybrid polypeptides interact.

In the selection methods described above, the activating domain can be p42 Gal 4, YP16 (HSV) and the DNA-binding domain can be derived from Gal4 or Lex A. The protein or enzyme can be adenylate cyclase, guanylate cyclase, DHFR and the like.

Examples of modulating compounds are set forth in columns 2 and 3 of Table II.

In yet another embodiment, the present invention relates to a pharmaceutical composition comprising the modulating compounds for preventing or treating Candida infection, cancer and neurodegenerative diseases in a human or animal, most preferably in a mammal.

This pharmaceutical composition comprises a pharmaceutically acceptable amount of the modulating compound. The pharmaceutically acceptable amount can be estimated from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes or encompasses a concentration point or range having the desired effect in an in vitro system. This information can thus be used to accurately determine the doses in other mammals, including humans and animals. The therapeutically effective dose refers to that amount of the compound that results in amelioration of symptoms in a patient. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or in experimental animals. For example, the LD50 (the dose lethal to 50% of the population) as well as the ED50 (the dose therapeutically effective in 50% of the population) can be determined using methods known in the art. The dose ratio between toxic and therapeutic effects is the therapeutic index which can be expressed as the ratio between LD 50 and ED50 compounds that exhibit high therapeutic indexes.

The data obtained from the cell culture and animal studies can be used in formulating a range of dosage of such compounds which lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity.

The pharmaceutical composition can be administered via any route such as locally, orally, systemically, intravenously, intramuscularly, mucosally, using a patch and can be encapsulated in liposomes, microparticles, microcapsules, and the like. The pharmaceutical composition can be embedded in liposomes or even encapsulated.

Any pharmaceutically acceptable carrier or adjuvant can be used in the pharmaceutical composition. The modulating compound will be preferably in a soluble form combined with a pharmaceutically acceptable carrier. The techniques for formulating and administering these compounds can be found in "Remington's Pharmaceutical Sciences" Mack Publication Co., Easton, PA, latest edition.

The mode of administration optimum dosages and galenic forms can be determined by the criteria known in the art taken into account the seriousness of the general condition of the mammal, the tolerance of the treatment and the side effects.

The present invention also relates to a method of treating or preventing Candida infection, cancer and neurodegenerative diseases in a human or mammal in need of such treatment. This method comprises administering to a mammal in need of such treatment a pharmaceutically effective amount of a modulating compound which binds to a targeted Candida protein or a protein involved in cancer or neurodegenerative diseases. In a preferred embodiment, the modulating compound is a polynucleotide which may be placed under the control of a regulatory sequence which is functional in the mammal or human.

In yet another embodiment, the present invention relates to a pharmaceutical composition comprising a SID® polypeptide, a fragment or variant thereof. The SID® polypeptide, fragment or variant thereof can be used in a pharmaceutical composition provided that it is endowed with highly specific binding properties to a bait polypeptide of interest. The original properties of the SID® polypeptide or variants thereof interfere with the naturally occurring interaction between a first protein and a second protein within the cells of the organism. Thus, the SID® polypeptide binds specifically to either the first polypeptide or the second polypeptide.

Therefore, the SID® polypeptides of the present invention or variants thereof interfere with protein-protein interactions between Candida proteins or between a Candida protein and a human or mammal protein.

Thus, the present invention relates to a pharmaceutical composition comprising a pharmaceutically acceptable amount of a SID® polypeptide or variant thereof, provided that the variant has the above-mentioned two characteristics; i.e., that it is endowed with highly specific binding properties to a bait polypeptide of interest and is devoid of biological activity of the naturally occurring protein.

In yet another embodiment, the present invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of a polynucleotide encoding a

SID® polypeptide or a variant thereof wherein the polynucleotide is placed under the control of an appropriate regulatory sequence. Appropriate regulatory sequences that are used are polynucleotide sequences derived from promoter elements and the like.

Polynucleotides that can be used in the pharmaceutical composition of the present invention include the nucleotide sequences of uneven SEQ ID from n°1 to 863 (column 2, Table II).

Besides the SID® polypeptides and polynucleotides, the pharmaceutical composition of the present invention can also include a recombinant expression vector comprising the polynucleotide encoding the SID® polypeptide, fragment or variant thereof.

The above described pharmaceutical compositions can be administered by any route such as orally, systemically, intravenously, intramuscularly, intradermally, mucosally, encapsulated, using a patch and the like. Any pharmaceutically acceptable carrier or adjuvant can be used in this pharmaceutical composition.

The SID® polypeptides as active ingredients will be preferably in a soluble form combined with a pharmaceutically acceptable carrier. The techniques for formulating and administering these compounds can be found in "Remington's Pharmaceutical Sciences" supra.

The amount of pharmaceutically acceptable SID® polypeptides can be determined as described above for the modulating compounds using cell culture and animal models.

Such compounds can be used in a pharmaceutical composition to treat or prevent Candida infection, cancer and neurodegenerative diseases.

Thus, the present invention also relates to a method of preventing or treating Candida infection, cancer and neurodegenerative diseases in a mammal said method comprising the steps of administering to a mammal in need of such treatment a pharmaceutically effective amount of: a SID® polypeptide of even SEQ ID from n°2 to 864 (column 3, Table II) or a variant thereof which binds to a targeted Candida, yeast, human or mammal protein; or or SID® polynucleotide encoding a SID® polypeptide of even SEQ ID from n^c 2 to 864 (column 3, Table II) or a variant or a fragment thereof wherein said polynucleotide is placed under the control of a regulatory sequence which is functional in said mammal; or a recombinant expression vector comprising a polynucleotide encoding a SID® polypeptide which binds to a yeast, Candida protein.

In another embodiment the present invention nucleic acids comprising a sequence of uneven SEQ ID from n° 1 to 863 (column 2, Table II) which encodes the protein of even SEQ ID from n° 2 to 864 (column 3, Table II) and/or functional derivatives thereof are administered to modulate complex (from Table I) function by way of gene therapy. Any of the methodologies relating to gene therapy available within the art may be used in the practice of the present invention such as those described by Goldspiel et al Clin. Pharm. 12 pgs. 488-505 (1993).

Delivery of the therapeutic nucleic acid into a patient may be direct in vivo gene therapy (i.e., the patient is directly exposed to the nucleic acid or nucleic acid-containing vector) or indirect ex vivo gene therapy (i.e., cells are first transformed with the nucleic acid in vitro and then transplanted into the patient).

For example for in vivo gene therapy, an expression vector containing the nucleic acid is administered in such a manner that it becomes intracellular; i.e., by infection using a defective or attenuated retroviral or other viral vectors as described, for example in U.S. Patent 4,980,286 or by Robbins et al, Pharmacol. Ther. , 80 No. 1 pgs. 35-47 (1998).

The various retroviral vectors that are known in the art are such as those described in Miller et al, Meth. Enzymol. 217 pgs. 581-599 (1993) which have been modified to delete those retroviral sequences which are not required for packaging of the viral genome and subsequent integration into host cell DNA. Also adenoviral vectors can be used which are advantageous due to their ability to infect non-dividing cells and such high-capacity adenoviral vectors are described in Kochanek, Human Gene Therapy, 10, pgs. 2451-2459 (1999). Chimeric viral vectors that can be used are those described by Reynolds et al, Molecular Medecine Today, pgs. 25 -31 (1999). Hybrid vectors can also be used and are described by Jacoby et al, Gene Therapy, 4, pgs. 1282-1283 (1997).

Direct injection of naked DNA or through the use of microparticle bombardment (e.g., Gene Gun®; Biolistic, Dupont). or by coating it with lipids can also be used in gene therapy. Cell-surface receptors/transfecting agents or through encapsulation in liposomes, microparticles or microcapsules or by administering the nucleic acid in linkage to a peptide which is known to enter the nucleus or by administering it in linkage to a ligand predisposed to receptor-mediated endocytosis (See, Wu & Wu, J. Biol. Chem., 262 pgs. 4429-4432 (1987)) can be used to target cell types which specifically express the receptors of interest. In another embodiment a nucleic acid ligand compound may be produced in which the ligand comprises a fusogenic viral peptide designed so as to disrupt endosomes, thus allowing the nucleic acid to avoid subsequent lysosomal degradation. The nucleic acid may be targeted in vivo for cell specific endocytosis and expression by targeting a specific receptor such as that described in WO92/06180, W093/14188 and WO 93/20221. Alternatively the nucleic acid may be introduced intracellularly and incorporated within the host cell genome for expression by homologous recombination. See, Zijlstra et al, Nature, 342, pgs. 435-428 (1989).

In ex vivo gene therapy, a gene is transferred into cells in vitro using tissue culture and the cells are delivered to the patient by various methods such as injecting subcutaneously, application of the cells into a skin graft and the intravenous injection of recombinant blood cells such as hematopoietic stem or progenitor cells.

Cells into which a nucleic acid can be introduced for the purposes of gene therapy include, for example, epithelial cells, endothelial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes and blood cells. The blood cells that can be used include, for example, T- lymphocytes, B-lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryotcytes, granulocytes, hematopoietic cells or progenitor cells and the like.

In yet another embodiment the present invention relates to protein chips or protein microarrays. It is well known in the art that microarrays can contain more than 10,000 spots of a protein that can be robotically deposited on a surface of a glass slide or nylon filter. The proteins attach covalently to the slide surface, yet retain their ability to interact with other proteins or small molecules in solution. In some instances the protein samples can be made to adhere to glass slides by coating the slides with an aldehyde-containing reagent that attaches to primary amines. A process for creating microarrays is described, for example by MacBeath and Schreiber in Science, Volume 289, Number 5485, pgs, 1760-1763 (2000) or Service, Science, Vol, 289, Number 5485 pg. 1673 (2000). An apparatus for controlling, dispensing and measuring small quantities of fluid is described, for example, in U.S. Patent No. 6,112,605.

The present invention also provides a record of protein-protein interactions, PIM®'s, SID®'s and any data encompassed in the following Tables. It will be appreciated that this record can be provided in paper or electronic or digital form. In order to fully illustrate the present invention and advantages thereof, the following specific examples are given, it being understood that the same are intended only as illustrative and in nowise limitative.

EXAMPLES Medium composition and standard protocols are available in Maniatis et al..

Example 1 : Preparation of a Saccharomyces cerevisiae genomic collection 1.A. Collection preparation and transformation in Eschenchia coli

1.A.1. Fragmented of genomic DNA preparation The Saccharomyces cerevisiae (strain YM955 (Matα, ura3-52, his3-200, ade2-101 ,

Iys2-801 , leu2-3, trp1-901 , tyr1-501 , gal4-542, gal80-538)) genomic DNA were fragmented in a nebulizer (GATC) for 1 minute, precipitated and resuspended in water.

The obtained nebulized genomic DNA was successively treated with Mung Bean Nuclease (Biolabs) (30 minutes at 30°C), T4 DNA polymerase (Biolabs) (10 minutes at 37°C) and Klenow enzyme (Pharmacia) (10 minutes at room temperature and 1 hour at 16°C). DNA was then extracted, precipitated and resuspended in water. 1.A.2. Ligation of linkers to blunt-ended genomic DNA

Oligonucleotide PL160 (5' end phosphorylated) 1 μg/μl and PL159 2μg/μl were used. Sequence of the oligo PL160 : 5'-ATCCCGGACGAAGGCC-3' (SEQ ID No.865) Sequence of the oligo PL159 : 5'-GGCCTTCGTCCGG-3'(SEQ ID No.866)

Linkers were preincubated (5 minutes at 95°C, 10 minutes at 68°C, 15 minutes at 42°C) then cooled down at room temperature and ligated with genomic DNA inserts at 4°C overnight.

Linkers were further removed on a separation column (Chromaspin TE 400, Clontech), according to the manufacturer protocol. 1.A.3. Vector preparation pP2 (Figure 8) was successively digested with BamHI restriction enzyme (Biolabs) for 1 hour at 37°C, dephosphorylated with Calf Intestine Phosphatase (CIP) (Biolabs) and filled in with dGTP using Vent DNA polymerase (exo-) (Biolabs), extracted, precipitated and resuspended in water.

1.A.4. Ligation between vector and insert of genomic DNA

The prepared vector was ligated overnight at 15°C with the genomic blunt ended DNA described in section 2 using T4 DNA ligase (Biolabs). The DNA was then precipitated and resuspended in water. 1-4.5. Library transformation in Escherichia coli

DNA from section 1.A.4 was then transformed into Electromax DH10B electrocompetent cells (Gibco BRL) with Cell Porator apparatus (Gibco BRL). 1 ml SOC medium was added and transformed cells were incubated at 37°C for 1 hour. 9 ml volume of

SOC medium per tube was added and plated on LB+ampicillin medium. Colonies were scraped with liquid LB medium, aliquoted and frozen at -80°C.

The obtained collection of recombinant cell clones is named HGXYeastB. 1.B. Collection transformation in Saccharomyces cerevisiae

The Saccharamyces cerevisiae strain (Y187 (MATα Gal4Δ GalδOΔ ade2-101 His3 Leu2-3, -1 12 Trp1-901 Ura3-52 URA3::UASGAL1-LacZ Met)) was transformed with the HGXYeastB Saccharomyces cerevisiae genomic DNA library. The plasmid DNA contained in E. coli was extracted (Qiagen) from aliquoted E. coli frozen cells (1.A.5.).

Saccharomyces cerevisiae yeast Y187 was grown in YPGIu.

Yeast transformation was performed according to standard protocol (Giest ef al. Yeast, 11 , 355-360, 1995) using yeast carrier DNA (Clontech). This experiment lead to 10⁴ to 5 x10⁴ cells/μg DNA. 2 x10⁴ cells were spread on DO-Leu medium per plates. Cells were aliquoted and frozen at -80°C.

The obtained collection of recombinant cell clones is named HGXYeastY. 1.C. Construction of bait plasmid

The genomic amplification of the ORF was obtained by PCR using the Pfu proofreading Taq polymerase (Stratagene) and 200 ng of genomic DNA as template. PCR primers were chosen in regions flanking the ORF. The PCR program was set up as follows : 94° 45"

94° 45¹

4488°° 4455"" x 30 cycles

7 722°° 66''

72° 10' 15° ∞ The amplification was checked on agarose gel. PCR fragments were purified with Qiaquick column (Qiagen) according to the manufacturer protocol.

Purified PCR fragments were digested with adequate restriction enzymes. Digested PCR fragments were purified with Qiaquick column (Qiagen) according to the manufacturer protocol.

Purified and digested PCR fragments were ligated into an adequately digested and dephosphorylated bait vector (pB6, Figure 3) according to standard protocol (Maniatis et al.).

Competent bacterial cells were transformed . The cells were grown, the DNA extracted and the plasmid was sequenced.

Example 2: Screening the collection with the two-hybrid in yeast system 2.A. The mating protocol

The mating two-hybrid in yeast system has been chosen (first described by Legrain et al., Nature Genetics, 1997, vol. 16, 277-282, Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens) for its advantages but the Saccharomyces cerevisiae collection could also have been screened in the classical two-hybrid system as described in Fields et al. or in a yeast reverse two-hybrid system.

The mating procedure allowed a direct selection on selective plates because the two fusion proteins were already produced in the parental cells. No replica plating is required.

This protocol was written for the use of the library transformed into the Y187 strain. Before mating, S. cerevisiae (CG 1945 strain (MATa Gal4-542 Gall 80-538 ade2-101 His3^*200 Leu2-3,-1 12 Trp1-901 Ura3-52 Lys2-801 URA3::GAL4 17mers (X3)-CyC1TATA-LacZ LYS2::GAL1 UAS-GAL1TATA-HIS3 CYH^R)) was transformed according to step 1.B. and spread on DO-Trp medium.

Day 1 , morning : preculture

Y187 cells carrying the bait plasmid obtained at step 1.C were precultured in 20 ml DO- Trp medium and grown at 30°C with vigorous agitation. Day 1 , late afternoon : culture The OD_600nrn of the DO-Trp preculture of Y187 cells carrying the bait plasmid was measured. The OD₆oonm must lie between 0.1 and 0.5 in order to correspond to a linear measurement.

150 ml DO-Trp at OD600nm 0.006/ml was inoculated and grown overnight at 30°C with vigorous agitation. Day 2 : mating medium and plates

5 YPGIu plates 50 ml tube with 30 ml DO-Leu-Trp-His 100 ml flask with 20 ml of YPGIu 75 DO-Leu-Trp-His plates

2 DO-Leu plates 2 DO-Trp plates 2 DO-Leu-Trp plates The OD600nm of the DO-Trp culture was measured. It should be around 1. For the mating, twice as many bait cells as library cells were used. To get a good mating efficiency, one must collect the cells at 10⁸ cells per cm².

The amount of bait culture (in ml) that makes up 80 OD600nm units for the mating with the yeast collection was estimated.

A vial containing the HGXYeastY library was thawed slowly on ice. The contents of the vial was added to 20 ml YPGIu. Cells were recovered at 30°C, under gentle agitation for 10 minutes. Mating

The 80 ODδOOnm units of bait culture was placed into a 250 ml flask. The HGXYeastY library culture was added to the bait culture. The mixture of diploids was transferred into 50 ml sterile tubes and centrifuged. The supernatant was discarded and the cells resuspend in YPGIu medium.

Cells were distributed in 400 μl samples in YPGIu plates with glass beads and spread by shaking the plates. The plates were incubated cells-up at 30°C for 4h30min. Collection of mated cells

The plates were washed and rinsed plates and collected cells were spread on DO-Leu- Trp-His+Tet plates. Day 4

Clones that were able to grow on DO-Leu-Trp-His+Tetracyclin were selected. This medium allows one to isolate diploid clones presenting an interaction.

The His+ colonies were counted on control plates. The number of His+ cell clones will define which protocol is to be processed: Upon 20 x10⁶ His+ colonies : if the number of His+ cell clones > 285 : then process overlay and then luminometry protocols on blue colonies (2.B and 2.C). if the number of His+ cell clones < 285 : process luminometry protocol (2.C). The following step leads to the selection of the strongest interaction.

2.B. The X-Gal overlay assay

The X-Gal overlay assay was performed directly on the selective medium plates after scoring the number of His* colonies. Materials A waterbath was set up. The water temperature should be 50°C.

0.5 M Na₂HP0₄ pH 7.5. 1.2% Bacto-agar. 2% X-Gal in DMF. The overlay mixture was 0.25 M Na₂HP0₄ pH7.5, 0.5% agar, 0.1% SDS, 7% DMF (LABOSI), 0.04% X-Gal (ICN). For each plate, 10 ml overlay mixture are needed. DO-leu-trp-his plates. Sterile toothpicks. Experiment

The temperature of the overlay mix should be between 45 and 50°C. The overlay-mix was poured over the plates in portions of 10 ml and collected when the top layer was settled. The plates were incubated overlay-up at 30°C. The time was noted and for blue colonies were checked for regularly. If no blue colony appeared, overnight incubation was performed. Using a pen the number of positives was marked.

The positive colonies were streaked on fresh DO-Leu-Trp-His plates with a sterile toothpick.

2.C. The luminometry assay

His+ colonies were grown overnight at 30°C in microtiter plates containing DO-Leu-Trp-

His+Tetracyclin medium with shaking. The day after, the overnight culture was diluted 15 times into a new microtiter plate containing the same medium, and incubated for 5 hours at 30°C with shaking. Samples were diluted 5 times and read OD₆₀o_nm- Another dilution was performed again to obtain between 10 000 and 75 000 yeast cells/well in 100 μl final volume.

Per well, 76 μl of One Step Yeast Lysis Buffer (Tropix), 20 μl Sapphirell Enhancer (Tropix), 4 μl Galacton Star (Tropix) was added and incubated 40 minutes at 30^CC. The β-Gal read-out (L) was measured using a Luminometer (Trilux, Wallach).

The value of OD₆₀onmxL was calculated and interacting preys having the highest values were selected. At this step of the protocol, the diploid cell clones presenting an interaction were isolated. The next step was aimed at identifying polypeptides involved in the selected interactions. Example 3: Identification of positive clones

3.A. PCR on yeast colonies

Introduction

PCR amplification of fragments of plasmid DNA directly on yeast colonies was a quick and efficient procedure to identify sequences cloned into this plasmid. It is directly derived from a published protocol (Wang H. et al., Analytical Biochemestry, 237, 145-146, 1996). However, it is not a standardized protocol; it varies from strain to strain, it was dependent of experimental conditions (number of cells, Taq polymerase source, etc). This protocol should be optimized to specific local conditions.

Materials For 1 well, the PCR mix composition was :

32.5 μl water,

5 μl 10X PCR buffer (Pharmacia),

1 μl dNTP IO mM,

0.5 μl Taq polymerase (5u/μl) (Pharmacia), 0.5 μl oligonucleotide ABS1 10 pmole/μl: S'-GCGTTTGGAATCACTACAGG-S' (SEQ ID

No.867) 0.5 μl oligonucleotide ABS2 10 pmole/μl: 5'-CACGATGCACGTTGAAGTG-3'.(SEQ ID No. 868)

1 N NaOH.

Experiment Positive colonies were grown overnight at 30°C on a 96 well cell culture cluster

(Costar), containing 150 μl DO-Leu-Trp-His+Tetracyclin with shaking. The cultures were resuspended and 100 μl was transferred immediately to a Thermowell 96 (Costar), then centrifuged for 5 minutes at 4000 rpm at room temperature. The supernatant was then removed and 5 μl NaOH was dispensed in each well and shaken for 1 minute.

The Thermowell was placed in the thermocycler (GeneAmp 9700, Perkin Elmer) for 5 minutes at 99.9°C and then 10 minutes at 4°C.

Into each well, the PCR mix was added shaken well. The PCR program was set up as follows :

94°C 3 minutes

94°C 30 seconds

53°C 1 minute 30 seconds x 35 cycles

72°C 3 minutes

72°C 5 minutes

15°C OO

The quality, the quantity and the length of the PCR fragment was checked on agarose gel.The length of the cloned fragment was the estimated length of the PCR fragment minus 300 base pairs that corresponded to the amplified flanking plasmid sequences. 3.B. Plasmids rescue from yeast by electroporation

Introduction

The previous protocol of PCR on yeast cell may not be successful, in such a case, plasmids may be rescued from yeast by electroporation. This experiment allows the recovery of prey plasmids from yeast cells by transformation of E. coli with a yeast cellular extract. One can then amplify the prey plasmid and sequence the cloned fragment.

Material

Plasmid rescue

Glass beads 425-600 μm (Sigma)

Phenol/chloroform (1/1 ) premixed with isoamyl alcohol (Amresco) Extraction buffer: 2% Triton X100, 1% SDS, 100 mM NaCl, 10 mM TrisHCI pH 8.0,

1 mM EDTA pH 8.0. Mix ethanol/NH^c: 6 volumes ethanol with 7.5 M NH₄ Acetate, 70% Ethanol and yeast cells in patches on plates.

Electroporation

SOC medium M9 medium

Selective plates : M9-Leu+Ampicillin

2 mm electroporation cuvettes (Eurogentech)

Experiment

Plasmid rescue The cell patch was prepared on DO-Leu-Trp-His with the cell culture of section 2.C.

The cell of each patch was scraped into an Eppendorf tube, 300 μl of glass beads was added in each tube, then, 200 μl extraction buffer was added and then 200 μl phenol:chloroform:isoamyl alcohol (25:24:1 ) was added. The tubes were centrifuged for 10 minutes at 15,000 rpm. 180 μl of supernatant was transferred to a sterile Eppendorf tube and 500 μl ethanol/NH^c, vortex was added to each tube. The tubes were centrifuged for 15 minutes, at 15,000 rpm at 4°C. The pellet was then washed with 200 μl 70% ethanol, then the ethanol was removed and the pellet was dried and resuspended in 10 μl water. The extracts were stored at -20°C. Electroporation

Electrocompetent MC1066 cells were prepared according to standard protocols (Maniatis). 1 μl of yeast plasmid DNA-extract was added to a pre-chilled Eppendorf tube, and kept on ice.

1 μl plasmid yeast DNA-extract sample was mixed and 20 μl electrocompetent cells were added and transferred into a cold electroporation cuvette. A Biorad electroporator was set on 200 ohms resistance, 25 μF capacity; 2.5 kVolts. The cuvette was placed in the cuvette holder and the sample was electroporated.

1 ml SOC was added into the cuvette and the cell-mix was transferred into a sterile Eppendorf tube. Cells were recovered for 30 minutes at 37°C and spun down for 1 minute, at 4000x g and the supernatant was poured off. About 100 μl medium was kept and used to resuspend the cells and spread them on selective plates (e.g., M9-Leu plates).

The plates were incubated for 36 hours at 37°C.

One colony was grown and the plasmids were extracted. The presence and size of insert was checked through enzymatic digestion and agarose gel. The insert was then sequenced.

Example 4: Protein-protein interaction For each bait, the previously protocol lead to the identification of prey polynucleotide sequences. In order to identify a protein-protein interaction, the obtained prey polypeptide sequence has to be characterized regarding the Saccharomyces cerevisiae genome.

This was accomplished with a software program names blastwun (available on the Internet site if the University of Washington: http://bioweb.pasteur.fr/seganal/interfaces/blastwu.html. this is a development version of software for gene and protein identification through similarity searches of protein and nucleotide sequence databases).

Blastwun program compares prey polynucleotide insert sequence (rescued from prey plasmid) with whole Saccharomyces cerevisiae genome (available on Stanford web site: http://genome-www.stanford.edu/Saccharomyces/). This comparison lead to prey polynucleotide localizations in S. cerevisiae genome, each localization having a score depending on the homology of sequence. For each prey polynucleotide, inventors considered the localization with the highest score and, if the insert sequence was included in and was in phase with an Open Reading Frame, inventors can identify one prey polypeptide interacting with one bait polypeptide.

See Table I : Protein-protein interactions in Saccharomyces cerevisiae.

Example 5 : Identification of SID®

By comparing and selecting the intersection of every isolated prey fragments obtained from example 3 and that were included in the same polypeptide, one can define the Selected Interacting Domain (SID®) as illustrated in Figure 15. The SID® were illustrated in Table II.

Example 6: Screening of modulating agent

One specific specific interaction is selected. A permeabilized yeast cell is transformed with plasmids containing bait polypeptide and prey polypeptide of the specific interaction. A top agar is plated containing transformed permeabilized yeast cells on square boxes

(that already contains agarose gel). The compounds to test are applied by spotting on top agar as soon as it is solidified, and incubated overnight at 30°C.

The results are then analysed and lead compounds that prevent transformed permeabilized yeast cells from growing are selected. Example 7: Making of polyclonal and monoclonal antibodies

The protein-protein complex of Table 1 is injected into mice and polyclonal and monoclonal antibodies are made following the procedure set forth in Sambrook et al supra.

More specifically mice are immunized with an immunogen comprising complexes conjugated to keyhole limpet hemocyanin using glutaraldehyde or EDC as is well known in the art. The complexes could also be stabilized by crosslinking as described in WO 00/37483. The immunogen is then mixed with an adjuvant. Each mouse receives four injections of 10 ug to

100 ug of immunogen, and after the fourth injection, blood samples are taken from the mice to determine if the serum contains antibodies to the immunogen. Serum titer is determined by ELISA or RIA. Mice with sera indicating the presence of antibody to the immunogen are selected for hybridoma production.

Spleens are removed from immune mice and single-cell suspension is prepared (Hariow et al 1988). Cell fusions are performed essentially as described by Kohler et al.. Briefly, P365.3 myeloma cells (ATTC Rockville, Md) or NS-1 myeloma cells are fused with spleen cells using polyethylene glycol as described by Hariow et al. Cells were plated at a density of 2 x 10⁵ cells/well in 96-well tissue culture plates. Individual wells are examined for growth and the supernatants of wells with growth are tested for the presence of Table I complex-specific antibodies by ELISA or RIA using Table I complex as a target protein. Cells in positive wells are expanded and subcloned to establish and confirm monoclonality.

Clones with the desired specificities are expanded and grown as ascites in mice or in a hollow fiber system to produce sufficient quantities of antibodies for characterization and assay development. Antibodies are tested for binding to bait polypeptide (from column 1 of Table I) alone or to prey polypeptide (from column 2 of Table I) alone, to determine which are specific for the Table I complex as opposed to those that bind to the individual proteins.

Monoclonal antibodies against each of the complexes set forth in Table I are prepared in a similar manner by mixing specified proteins together, immunizing an animal, fusing spleen cells with myeloma cells and isolating clones which produce antibodies specific for the protein complex, but not for individual proteins.

The following results obtained from these Examples, as well as the teachings in the specification are set forth in the Tables I and II below.

Table I depicts Saccharomyces cerevisiae interacting proteins.

Table II provides polynucleotide (column 2) and polypeptide (column 3) sequences of SID® interacting with a given bait (column 1 ).

While the invention has been described in terms of the various preferred embodiments, the skilled artisan will appreciate that various modifications, substitutions, omissions and changes may be made without departing from the scope thereof.

Table II - SID® sequences interacting with a given bait

YOROβOw 15 CATGAAGTACCATTTGGAAACATTACTTTACCAGCCGACTCAGAAGCTAGGAAAGCT 16 HEVPFGNITLPADSEARKAAIKFI GCAATAAAATTTATCAAATTCATCAATCCAAAGATTAATGATGGACAAATTCGCCATAT KFINPKINDGQIRHIPVRVYKNGL TCCAGTAAGGGTCTATAAGAACGGGCTTTGTGATGTTCCTCATATCCTAAAAGACAT CDVPHILKDIKYGKNSGEKLVAV CAAATATGGTAAGAACTCTGGTGAAAAACTCGTTGCCGTATTAAACTAGATGACGAA LN^*MTN RYYFLSYT*LLRLVIVT*F

CAGATATTATTTTCTTTCATATACATAGTTATTACGTTTAGTTATAGTGACTTAG I I I I I FFFF"NCLLLITPCRGNR*AQ*K

TTTTTTTTTTTAATGAAATTGTTTGTTATTGATCACCCCTTGTAGGGGCAACAGATGAG S^*LIM*FWSNPSMIIYPNYLLSNY

CACAGTGAAAATCATAACTCATTATGTGATTCGTTGTGAGTAACCCTTCCATGATTAT YISIELICTKHINLCTHFHWQRLG

ATATCCTAACTATCTACTATCCAACTACTACATATCTATTGAACTCATATGTACCAAGC EI^*YTVIISGEAL*VFLPIMAARCP

ACATAAATTTATGCACCCATTTCCATTGGCAAAGACTTGGCGAAATTTAATACACTGT NDCAALFCLLHSFFLRR^*AG*ME

AATAATATCCGGCGAAGCTCTTTAAGTATTCTTACCTATAATGGCGGCTCGCTGTCC TTIYCNRASPRNSHFPWIITKRV

GAACGATTGCGCCGCACTTTTCTGCCTCTTACATTCCTTTTTCCTAAGGCGATAAGCA LPFRSFSQQIQICRTCFRTTNRN

GGCTGAATGGAAACTACAATATATTGCAATAGAGCTTCCCCAAGGAACTCCCACTTT DVLQSTV*TESSRNILIMFFFFKL

CCTTGGATTATTACAAAGAGAGTTTTACCTTTTAGGTCATTCTCACAGCAGATCCAAA ANVNRLECMGTHHFVTDSIFYL

TATGTAGAACCTGTTTTAGAACAACAAATAGAAATGATGTGTTACAGAGTACCGTTTA P^*L*KRLSIV^*KYAIHILSSSVNDK

AACTGAGTCATCACGTAATATTCTAATTATG I I I I I I I I I I I TAAATTAGCTAATGTAAA ALFMLMYMGCQTTLPKKTAQNI

CAGGCTGGAATGTATGGGAACTCATCACTTCGTAACTGATTCCATCTTCTATCTACCA YLCRAIL"YIGFHLMEALAKITFFI

TAACTTTAAAAAAGATTATCGATAGTCTAAAAATACGCGATACACATTCTTTCTTCGTC RLLVGKKLA*SQVQCCYYVGA*S

CGTT.AATGATAAAGCTCTTTTTATGTTAATGTACATGGGTTGTCAAACTACCCTTCCC ADHTLYLNSSYYNAHD GPSEF

AAAAAGACAGCACAAAATATATATTTATGCCGTGCTATTCTATGATAATATATTGGCTT YLFSPRQYYANFNQA

TCACCTAATGGAAGCCCTAGCGAAGATAACCTTTTTCATACGACTTTTGGTGGGAAA

GAAATTGGCATAGTCCCAAGTACAATGTTGTTATTATGTAGGAGCTTAATCCGCAGAT

CATACACTCTATCTCAATTCTTCGTATTATAACGCTCATGATTAATTAGGTCCTTCTGA

GTTCTATCTTTTCTCTCCGAGACAATATTATGCAAATTTTAACCAAGCAG

[ YHR040W 19 AGCACACCACTGCATTCACCTGCGTTAAGAAAAATGAGCTCGAGGGACAACGACGA 20 STPLHSPALRKMSSRDNDDSG

TAGCGGAGACAACGTCAACGGTAGAGGCACCTCACCTATCCCTAATCTAAACATAGA DNVNGRGTSPIPNLNIDKPSPSA

CAAGCCTTCGCCGTCAGCGTCGTCAGCGTCAAAAAGAGAATATTTAAGTGCATATCC SSASKREYLSAYPTLAHRDSSS

AACACTAGCTCACAGAGATTCATCATCTTCACTTAGTCCGCGGGGCAAAGGACAACG SLSPRGKGQRSSSSSSSSQRIY

GTCTTCATCGTCTTCCAGTTCCAGTCAAAGAATATATGTTTCCCCACCATCTCCCACA VSPPSPTGDFVHGSCADGDNG

GGTGATTTCGTACACGGGAGTTGTGCAGACGGTGATAATGGATCTAGGACTAATACT SRTNTMVEMKRKKPVRPVDIDE

ATGGTCGAAATGAAAAGGAAAAAACCAGTTCGTCCAGTGGACATAGATGAAATTATC IIQRLLDAGYAAKRTKNVCLKNS

CAGAGATTACTAGATGCCGGCTATGCCGCAAAGAGGACCAAGAATGTTTGCTTAAAG EIIQICHKARELFLAQPALLELSP

AATTCCGAGATTATTCAAATTTGCCATAAGGCTCGTGAATTATTCCTTGCCCAACCTG SVKIVGDVHGQYADLLRLFTKC

CTCTCTTAGAATTATCTCCCTCGGTAAAAATAGTGGGTGATGTTCACGGCCAATATG GFPPMANYLFLGDYVDRGKQSL

CAGATCTTTTGAGACTTTTTACCAAATGCGGTTTCCCCCCCATGGCAAATTACTTATT ETILLLLCYKIKYPENFFLLRGNH

TTTAGGCGATTACGTAGATCGCGGTAAGCAGTCCCTGGAGACCATTTTACTATTACT ECANVTRVYGFYDECKRRCNIKI

ATGCTATAAGATTAAATATCCTGAAAATTTCTTCCTCTTAAGAGGCAATCATGAATGT WKTFVDTFNTLPLAAIVTGKIFC

GCCAATGTTACAAGAGTCTACGGGTTTTATGATGAATGTAAACGACGTTGTAATATCA VHGGLSPVLNSMDEIRHVSRPT

AGATTTGGAAAACCTTTGTTGACACGTTCAACACGCTACCGTTAGCAGCCATCGTCA DVPDFGLINDLLWSDPTDSSNE

CAGGAAAAATATTTTGTGTTCATGGTGGACTATCACCTGTTCTAAATTCCATGGACGA WEDNERGVSFCYNKVAINKFLN

AATTAGGCACGTTAGTAGGCCCACCGATGTACCCGACTTCGGCTTAATTAATGACCT KFGFDLVCRAHMWEDGYEFFN

TTTATGGTCGGATCCTACAGATTCATCGAATGAATGGGAGGATAATGAGCGTGGAGT DRSLVTVFSAPNYCGEFDNWG

TAG I I I I I GTTACAATAAAGTGGCTATTAATAAA I I I I I AAACAAATTCGGATTCGATT AVMTVSEGLLCSFELLDPLDSTA

TAGTGTGTAGAGCACATATGGTGGTGGAAGATGGTTATGAATTCTTTAATGACAGAA LKQVMKKGRQERKLANR'KTRY

GCTTAGTTACAGTGTTTTCGGCTCCCAACTATTGTGGTGAATTCGATAACTGGGGTG *CCS"AH*KLSLWPLP

CTGTCATGACCGTTAGTGAAGGCCTACTCTGTTCTTTTGAGTTGTTGGACCCACTGG

ACAGTACCGCTTTGAAACAAGTGATGAAAAAAGGCAGGCAAGAACGTAAATTAGCCA

ATCGCTGAAAAACTAGATATTAATGTTGCAGTTAATAAGCTCATTGAAAACTTTCTTTG

GTTGTTCCTCTTCCTTC

YDR530C 1 75 | CGTCGTTGGTTCGGTAACACAAGAGTGATATCTCAGGACGCACTGCAGCATTTCAGA 76 RRWFGNTRVISQDALQHFRSAL

AGTGCATTGGGTGAAACCCAAAAAGATACATATCAAGTTCTTCTGAGAAGAAATAAG GETQKDTYQVLLRRNKLPMSLL CTTCCCATGTCATTGTTGGAAGAAAAGGACGCAGATGAATCCCCAAAAGCCAGAATT EEKDADESPKARILDTESYADAF TTGGATACCGAAAGTTATGCTGATGCGTTTGGGCCCAAAGCCCAAAGAAAGAGACC GPKAQRKRPRLAASNLEDLVKA

ACGTCTTGCTGCATCCAATCTAGAGGACTTGGTCAAGGCTACAAATGAAGACATTAC TNEDITKYEEKQVLDATLGLMG CAAGTATGAGGAAAAGCAAGTCTTAGATGCCACATTAGGACTAATGGGGAACCAGG NQEDKENGWTSAAKEAIFSKGQ AAGACAAAGAAAATGGGTGGACCTCCGCAGCAAAAGAAGCTATTTTCAGTAAGGGTC SKRIWNELYKVIDSSDWIHVLD AATCCAAACGTATTTGGAACGAATTATATAAAGTCATCGATTCGTCTGACGTGGTAAT ARDPLGTRCKSVEEYMKKETPH ACATGTTCTAGATGCAAGGGATCCATTGGGTACCCGTTGTAAGTCCGTGGAAGAGTA KHLIYVLNKCDLVPTWVAVC*LF TATGAAGAAGGAAACACCACACAAGCATTTAATTTATGTTCTTAATAAGTGTGATTTG FFFF*FFPTKRIAREIFFFDRV^*FL GTACCTACCTGGGTTGCAGTATGTTAAC I I I I I I I I I I I I I I I I I I GATTCTTTCCAAC LPNLFVRVLLYPF*DKLIYRSVCY CAAGAGGATAGCACGAGAGATTTTTTTTTTCGATAGAGTATGATTTTTATTACCAAAC CFGRVILFAYPYFPVLGKPHG^*I CTTTTTGTCAGGGTGCTTCTCTATCCGTTTTAGGATAAACTTATCTACAGAAGTGTCT RRTTFVLKVYLRFLEQRGSYEL GTTACTGTTTTGGAAGAGTAATACTGTTCGCTTATCCATATTTCCCCGTTCTGGGGAA SFCYLRLYLGLHTICS"RSSQFS GCCTCATGGGTAAATTAGAAGGACAACCTTTGTTTTAAAGGTATACCTTCGCTTTTTA TFFFFLV^*STLRNNQSRL^*Q^*LLH GAACAGCGAGGATCTTATGAGTTGAGCTTTTGTTATTTGAGACTTTATCTCGGGCTC SLGAKDYLL^*NKMYFLLTEGFFF CATACAATATGTTCGTAGTAAAGATCCTCACAATTTTCTAC I I I I I I I I I I I I I I I AGTT QF*LYK^*AAWVKHLSKERPTLAF TAATCCACTCTTCGAAACAATCAATCTCGGCTATAACAATGATTATTACATTCTCTTGG HASITNSFGKGSLIQLLRQFSQL GGCAA GATTACCTATTATGAAACAAAATGTATTTTTTACTAACAGAAGGTTTTTTTT HTDRKQISVGFIGYPNTGKSSIIN TTCAATTCTAACTTTACAAGTAGGCAGCTTGGGTCAAACATTTGTCAAAGGAACGTCC TLRKKKVCQVAPIPG AACTTTGGCGTTTCACGCATCTATTACCAACTCTTTCGGTAAAGGTTCGTTAATTCAG TTGCTACGTCAGTTCTCACAGTTGCATACTGATAGAAAGCAAATATCTGTAGGGTTTA TCGGCTATCCAAACACTGGTAAATCGTCCATCATTAACACATTGAGAAAGAAAAAGG TGTGTCAAGTTGCACCAATCCCTGGTGA

NER107C 105 ATGGCAAGCATCGGTTCGCAAGTGAGAAAAGCTGCTTCTAGTATTGACCCTATCGTC 106 MASIGSQVRKAASSIDPIVTDYA

ACGGATTACGCAGTGGGCTACTTTAACCACTTGTCCGGAATAACTTTTGATGCTGTT VGYFNHLSGITFDAVQSKQVDL

CAAAGTAAGCAGGTAGATTTGTCCACTGAAGTGCAATTTGTGTCCGATTTATTGATTG STEVQFVSDLLIDAGASKAKVKE

ATGCGGGTGCGTCAAAGGCTAAAGTTAAAGAACTATCGGAAAGTATTTTGAAGCAAT LSESILKQLTTQLKENEAKLELT

TGACTACTCAACTAAAGGAGAACGAAGCCAAATTGGAATTGACCGGTGATACGTCCA GDTSKRLLDINVLKSHNSKSDIN

AGAGATTACTTGATATTAATGTCTTAAAGAGTCATAACAGTAAATCCGATATCAACGT VSLSMLGVNGDIEHTGRKMETR

CTCATTAAGCATGCTGGGTGTGAACGGTGACATCGAACATACTGGTAGAAAGATGGA VDLKKLAKAEQKIAKKVAKRNNK

AACAAGAGTTGATTTGAAAAAACTGGCCAAGGCTGAACAAAAGATCGCAAAGAAAGT FVKYEASKLINDQKEEDYDSFFL

CGCCAAGAGAAATAACAAATTTGTTAAATACGAGGCTTCTAAATTGATCAATGACCAA QINPLEFGSSAGKSKDIHIDTFDL

AAGGAGGAGGATTACGATTCTTTCTTTTTGCAAATCAACCCTTTAGAATTCGGTTCAT YVGDGQRILSNAQLTLSFGHRY

CCGCTGGTAAATCCAAGGATATCCATATTGACACTTTCGACTTGTACGTTGGTGACG GLVGQNGIGKSTLLRALSRREL

GTCAAAGAATTTTGTCCAACGCCCAATTGACTCTAAGTTTTGGTCACAGATATGGTCT NVPKHVSILHVEQELRGDDTKA

TGTGGGCCAAAATGGTATTGGTAAATCTACTTTGTTAAGGGCTCTATCTAGAAGAGA LQSVLDADVWRKQLLSEEAKIN

GCTGAACGTCCCCAAACATGTTTCGATTTTACACGTGGAACAAGAGTTAAGAGGTGA ERLKEMDVLRQEFEEDSLEVKK

TGATACAAAGGCTTTACAAAGTGTGCTGGATGCAGACGTTTGGAGAAAACAACTATT LDNEREDLDNHLIQISDKLVDME

AAGTGAAGAAGCCAAGATCAATGAAAGATTAAAGGAAATGGATGTATTAAGACAGGA SDKAEARAASILYGLGFSTEAQ

ATTCGAAGAAGACAGTTTAGAAGTTAAAAAATTGGACAATGAAAGAGAAGACTTGGA QQPTNSFSGGWRMRLSLARAL

TAACCATTTGATACAGATTTCTGACAAATTAGTCGATATGGAATCTGACAAGGCTGAA FCQPDLLLLDEPSNMLDVPSIAY

GCTAGGGCAGCATCAATCTTATATGGTTTGGGGTTCAGTACGGAGGCACAGCAACA LAEYLKTYPNTVLTVSHDRAFLN

ACCCACTAATTCCTTTTCCGGTGGTTGGAGAATGAGATTGTCCTTGGCAAGAGCCTT EVATDIIYQHNERLDYYRGQDF

ATTCTGTCAACCAGATCTTTTGTTGTTAGATGAACCTTCCAATATGTTGGATGTGCCA DTFYTTKEERRKNAQREYDNQ

TCCATCGCTTATTTAGCAGAGTATTTGAAAACATATCCAMTACAGTTTTGACAGTTTC MVYRKHLQEFIDKYRYNAAKSQ

TCACGACCGTGCATTCTTGAATGAAGTGGCTACAGATATCATTTATCAACACAACGAA EAQSRIKKLEKLPVLEPPEQDKT

AGACTAGACTATTACAGAGGCCAAGATTTCGATACCTTTTACACCACAAAGGAGGAA IDFKFPECDKLSPPIIQLQDVSFG

CGTAGAAAGAATGCTCAACGTGAGTATGATAACCAAATGGTTTACAGAAAGCACTTG

YNL230C 121 CAAGAAGGGGGCTTTATCCGTAGAAGGCGTACGAGAAGTACGAAAAAAAGTGTTAAT 122 QEGGFIRRRRTRSTKKSVNYNE

TATAATGAACTAAGCGATGACGATACAGCTGTGAAAAACTCAAAAACTCTGCAGCTG LSDDDTAVKNSKTLQLKGNSEN

AAAGGGAACAGCGAAAACGTAAATGACAGCCAAGATGAAGAATACCGTGATGATGC VNDSQDEEYRDDATLVKSPDD

TACGCTCGTGAAATCACCCGACGACGATGACAAAGATTTTATCATAGACCTAACAGG DDKDFIIDLTGSDKERTATDENT

TTCAGATAAAGAGCGGACCGCCACCGATGAGAATACTCATGCGATCAAGAATGATAA HAIKNDNDEIIEIKEERDVSDDDE

TGACGAGATAATAGAAATCAAGGAGGAACGTGATGTTTCGGATGACGACGAACCGTT PLTKKRKTTARKKKKKTSTKKKS

AACAAAGAAAAGGAAGACAACTGCTAGGAAGAAGAAGAAAAAAACGAGCACCAAGA PKVTPYERNTLRLYEHHPELRN

AAAAGTCACCGAAGGTAACCCCATATGAAAGAAACACTTTGCGATTATATGAGCATC VFTDLKNAPPYVPQRSKQPDG

ATCCTGAACTAAGGAATGTTTTCACGGATTTGAAAAATGCACCTCCCTATGTCCCCCA MTIKLLPFQLEGLHWLISQEESIY

AAGATCCAAGCAGCCGGATGGTATGACCATCAAACTGCTACCTTTCCAGTTAGAAGG AGGVLADEMGMGKTIQTIALLM

TCTTCATTGGCTAATATCTCAAGAGGAGAGCATTTATGCGGGCGGTGTTTTGGCAGA NDLTKSPSLWAPTVALMQWKN

CGAAATGGGTATGGGTAAGACCATCCAAACTATTGCCCTATTAATGAACGATTTGAC EIEQHTKGQLKIYIYHGASRTTDI

TAAGTCTCCGTCTTTAGTTGTTGCCCCTACCGTGGCGCTGATGCAGTGGAAAAACGA KDLQGYDWLTTYAVLESVFRK

AATAGAACAACATACAAAGGGACAACTGAAAATATACATTTATCACGGTGCTTCCAGA QNYGFRRKNGLFKQPSVLHNID

ACCACGGATATCAAAGATTTGCAAGGCTACGATGTTGTACTAACCACTTACGCAGTG FYRVILDEAHNIKDRQSNTARAV

CTGGAATCGGTATTCAGAAAGCAAAACTACGGGTTTAGAAGGAAAAATGGACTTTTC NNLKTQKRWCLSGTPLQNRIGE

AAGCAGCCTTCCGTATTGCATAATATTGACTTTTATAGAGTTATTCTGGATGAGGCAC M YSLI RFLN INPFTKYFCTKCDC

ACAATATCAAGGATAGACAAAGCAATACTGCTAGGGCTGTAAACAACTTAAAAACGC ASKDWKFTDRMHCDHCSHVIM

AAAAGCGATGGTGCCTGTCGGGTACTCCGCTGCAAAATAGAATTGGTGAGATGTATT QHTNFFNHFMLKNIQKFGVEGP

CTTTGATCAGATTCTTAAATATCAATCCTTTCACAAAGTACTTTTGTACCAAGTGTGAT GLESFNNIQTLLKNIMLRRTKVE

TGCGCTTCGAAGGACTGGAAATTTACGGATCGCATGCATTGTGACCATTGTAGTCAC RADDLGLPPRIVTVRRDFFNEE

GTCATTATGCAACACACGAATTTCTTCAACCATTTCATGTTGAAGAACATTCAGAAAT EKDLYRSLYTDSKRKYNSFVEE

TTGGTGTGGAAGGTCCTGGTTTAGAGTCTTTTAATAACATTCAGACATTATTGAAAAA GWLNNYANIFTLITRMRQLADH

CATCATGCTGCGAAGAACTAAAGTGGAAAGAGCGGATGACTTGGGTCTACCGCCCA PDLVLKRLNNFPGDDIGWICQL

GAATTGTTACCGTGAGGAGAGACTTCTTCAATGAAGAGGAAAAAGATCTTTACAGAA CNDEAEEPIESKCHHKFCRLCIK

YNL230C 141 GAAGAAGAGAAAGAAGCTAGGCATACAAATTCACCAAGGAAAAGAGGCCGTAAGAT 142 EEEKEARHTNSPRKRGRKIKLG

AAAACTAGGTAAAGATGATATTGACGCTTCTGTACAACCTCCCCCCAAAAAAAGAGG KDDIDASVQPPPKKRGRKPKDP

TCGTAAACCTAAAGATCCTAGTAAACCGCGTCAGATGCTATTGATATCTTCATGCCGT SKPRQMLLISSCRANNTPVIRKF

GCAAATAATACTCCTGTGATTAGGAAATTTACAAAAAAGAATGTTGCTAGGGCGAAAA TKKNVARAKKKYTPFSKRFKSIA

AGAAATATACCCCGTTTTCGAAAAGATTTAAATCTATAGCTGCAATACCAGATTTAAC AIPDLTSLPEFYGNSSELMASRF

TTCATTACCTGAATTTTACGGAAATTCTTCGGAATTGATGGCATCAAGGTTTGAAAAC ENKLKTTQKHQIVETIFSKVKKQ

AAATTAAAAACAACCCAAAAGCATCAGATTGTAGAAACAATTTTTTCTAAAGTCAAAAA LNSSYVKEEILKSANFQDYLPAR

ACAGTTGAACTCTTCGTATGTCAAAGAAGAAATATTGAAGTCTGCAAATTTCCAAGAT ENEFASIYLSAYSAIESDSATTIY

TATTTACCGGCTAGGGAGAATGAATTCGCCTCAATTTATTTAAGTGCATATAGTGCCA VAGTPGVGKTLTVREWKELLS

TTGAGTCCGACTCCGCTACTACTATATACGTGGCTGGTACGCCTGGTGTAGGGAAAA SSAQREIPDFLYVEINGLKMVKP

CTTTAACCGTAAGGGAAGTCGTAAAGGAACTACTATCGTCTTCTGCACAACGAGAAA TDCYETLWNKVSGERLTWAAS

TACCAGACTTTCTTTATGTGGAAATAAATGGATTGAAAATGGTAAAACCCACAGACTG MESLEFYFKRVPKNKKKTIWLL

TTACGAAACTTTATGGAACAAAGTGTCAGGAGAAAGGTTAACATGGGCAGCTTCAAT DELDAMVTKSQDIMYNFFNWTT

GGAGTCACTAGAGTTTTACTTTAAAAGAGTTCCAAAAAATAAGAAGAAAACCATTGTA YENAKLIVIAVANTMDLPERQLG

GTCTTGTTGGACGAACTCGATGCCATGGTAACGAAATCTCAAGATATTATGTACAATT NKITSRIGFTRIMFTGYTHEELKN

TTTTCAATTGGACTACTTACGAAAATGCCAAACTTATTGTCATTGCAGTAGCCAATAC IIDLRLKGLNDSFFYVDTKTGNAI

AATGGACTTACCAGAACGTCAGCTAGGCAATAAGATTACTTCAAGAATTGGGTTTAC LIDAAGNDTTVKQTLPEDVRKV

CAGAATTATGTTCACTGGGTATACGCACGAAGAGCTAAAAAATATCATTGATTTAAGA RLRMSADAIEIASRKVASVSGDA

CTGAAGGGGTTGAACGACTCATTTTTCTATGTTGATACAAAAACTGGCAATGCTATTT RRALKVCKRAAEIAEKHYMAKH

TGATTGATGCGGCTGGAAACGACACTACAGTTAAGCAAACGTTGCCTGAAGACGTG GYGYDGKTVIEDENEEQIYDDE

AGGAAAGTTCGCTTAAGAATGAGTGCTGATGCCATTGAAATAGCTTCGAGAAAAGTA DKDLIESNKAKDDNDDDDDNDG

GCAAGTGTTAGTGGTGATGCAAGAAGAGCATTGAAGGTTTGTAAAAGAGCAGCTGAA VQTVHITHVMKALNETLNSHVIT

ATTGCTGAAAAACACTATATGGCTAAGCATGGTTATGGATATGATGGAAAGACGGTT FMTRLSFTAKLFIYALLNLMKKN

ATTGAAGATGAAAATGAGGAGCAAATATACGATGATGAAGACAAGGATCTTATTGAA GSQEQELGD

YNL286W 157 CAGCCAAATAAGGGAACTAGCAGGTGGACAATTGGATCTGATACTGAATCTTCTAGA 158 QPNKGTSRWTIGSDTESSREPS

GAACCTAGTATTAGCCCAAACGAGAACACAACTTCAATTACTAAATCGCGAAACCGC ISPNENTTSITKSRNRNKKASKP

AATAAGAAGGCTAGCAAGCCGACGCTGAATGAAAAATCAAAAACAACAACGATGCCA TLN EKSKTTTMPKKLETKNQEK

AAAAAGTTGGAGACAAAAAATCAAGAGAAAAATAATGGAAAAACTAAGGACGGGAAA NNGKTKDGKLIYEEGEPLTRYN

CTTATTTACGAAGAAGGTGAACCACTCACAAGGTATAATACCTTTAAGTCAACCTTAT TFKSTLSYPDLNTYLNDYSFALF

CATACCCCGATTTGAACACATACCTAAATGATTACTCATTTGCCCTTTTTTTGGAACA LEQKLENEFVQNFNILWPRNEK

GAAATTAGAGAACGAATTTGTTCAAAACTTCAACATACTATGGCCTCGCAATGAAAAA DTAFIINVEKNNNSELEKLLPANL

GATACAGCTTTTATAATCAACGTTGAGAAGAATAATAATTCAGAACTTGAGAAATTAC LALGRPAFNERQPFFFCTQDEQ

TACCTGCAAATTTGCTTGCACTCGGTAGACCGGCATTTAATGAAAGGCAACCATTTTT KVWYIFIKELSIQRGKWLLVELF

CTTTTGTACTCAGGATGAACAAAAAGTTTGGTATATTTTTATCAAAGAACTTTCTATTC SWNNLSLPTKNGSSQFKLLPTS

AAAGGGGTAAATACGTGTTGTTAGTGGAGC I I I I I I CGTGGAATAATTTGAGTTTGCC AQTSRILFAMTRITNPKFIDLLLG

AACGAAAAATGGTTCATCTCAATTTAAGCTATTGCCAACTTCTGCGCAGACAAGTAGA QKPIKEIYFDNRLKFSSDKLNRS

ATTCTATTTGCCATGACGCGTATTACAAACCCAAAATTTATTGACCTATTACTAGGCC QKTAVEHVLNNSITILQGPPGTG

AAAAGCCTATTAAAGAGATTTATTTTGATAACAGGCTAAAATTTTCCAGTGACAAGTT KTSTIEEIIIQVIERFHAFPILCVAA

GAACCGATCTCAGAAGACTGCAGTGGAACATGTTCTAAACAACAGTATCACAATTCT SNIAIDNIAEKIMENRPQIKILRILS

ACAAGGCCCACCTGGTACAGGTAAAACATCAACAATAGAGGAGATAATCATCCAAGT KKKEQQYSDDHPLGEICLHNIVY

AATTGAAAGATTTCACGCATTCCCAATATTGTGTGTTGCCGCGTCAAATATTGCTATT KNLSPDMQWANKTRRGEMISK

GATAACATTGCCGAGAAGATTATGGAGAATAGACCGCAGATAAAGATCTTAAGAATT SEDTKFYKEKNRVTNKWSQSQ

TTGTCAAAGAAGAAAGAACAACAGTATAGTGATGACCACCCATTGGGCGAAATTTGT IIFTTNIAAGGRELKVIKECPWIM

CTCCACAATATTGTATACAAAAATCTTTCCCCAGACATGCAAGTCGTGGCAAACAAAA DEATQSSEASTLVPLS

CCCGCAGGGGCGAAATGATCTCCAAATCAGAGGACACGAAGTTTTATAAAGAGAAAA

ACCGTGTCACCAACAAGGTCGTATCTCAATCGCAAATAATTTTCACGACAAACATTGC

TGCAGGTGGTCGTGAATTGAAGGTTATAAAAGAATGTCCCGTGGTAATAATGGATGA

GGCTACACAATCATCAGAGGCTTCAACACTAGTCCCACTTTCGTT

YNL286W 203 TCTTACATCTCAACCGTTGAGAAAAATAATGATAGGAAGAACCTAACCGTAATAATTA 204 SYISTVEKNNDRKNLTVIITLEIN

CATTAGAAATCAATGGCCATCTAAGAGTTTTTACTATTCCAGATTTCAAAGAACAAAT GHLRVFTIPDFKEQMSEHIPFPI

GTCTGAACATATTCCATTTCCAATTGCCGCAAAATACATTACAGAATCATCTGTTTTAA AAKYITESSVLRNGDIAIRVSEFQ

GAAATGGTGATATAGCGATTCGAGTGAGTGAATTTCAGGCGTCACTATTTTCTACTGT ASLFSTVKEQDTLAPVSDTLYIN

CAAGGAACAGGATACGCTAGCGCCAGTTTCTGACACACTGTACATCAATGGAATCAG GIRIPYRPQVNSLQWARGTVYC

GATTCCCTATAGGCCACAAGTAAATTCATTACAGTGGGCAAGAGGCACTGTATATTG TPAQLNELLGGVNRPASKYKESI

TACTCCCGCTCAATTAAATGAATTATTGGGTGGCGTGAATAGACCTGCATCCAAATA IAEGSFSERSSDDNNANHPEHQ

CAAAGAGAGCATCATTGCTGAGGGAAGTTTCTCGGAGCGTTCTTCAGACGATAATAA YTKPTRKGRNSSYGVLRNVSRA

TGCGAACCATCCGGAACACCAGTATACCAAGCCCACCCGTAAGGGCAGGAACAGTA VETRWDAVEDRFNDYATAMGE

GTTACGGGGTACTAAGAAATGTTTCTCGAGCTGTAGAAACTAGATGGGACGCCGTTG TMNEAVEQTGKDVMKGALGF*I

AAGATAGATTCAACGACTATGCTACCGCCATGGGAGAAACAATGAACGAGGCTGTTG HTLIHTG"ATSFYTYFFLVLYAL*

AGCAGACTGGAAAAGATGTAATGAAAGGTGCCCTTGGTTTTTAAATACATACACTAAT INLNRFIFHYTCLVLRRFVNT^*AIK

TCATACAGGGTAATGAGCAACTTCCTTCTATACTTATT I I I I I I I AGTTCTATATGCTT FIIL^*LNE^*PHLDHLHEFVITFLEQ*

TATAAATTAATTTGAATCGATTTATTTTTCATTATACATGTCTAGTTTTACGAAGGTTTG EQCQPH*PIHHHHRQILGPIF^*HV

TAAATACATGAGCTATCAAGTTTATAATCCTTTAATTAAACGAGTGACCTCATTTAGAC IYLLSTPPYRGLRQCDNPLFQQ

CATCTTCATGAATTTGTGATAACTTTTCTTGAACAATAGGAACAATGTCAGCCGCACT HRIAS^*D^*PWPYRHIQLQCQHPII

AACCAATTCACCATCATCATCGGCAAATTCTTGGCCCAATCTTTTGACACGTAATTTA A"PSFFRVNNQSSPYHHQKP^*R

CCTTCTAAGTACTCCTCCTTACCGAGGATTACGGCAATGTGACAACCCGCTTTTTCA TPV^*PLAL*L*RN*FFV*SKKIFFQ

GCAGCATCGAATTGCTTCCTAGGATTAGCCTTGGCCTTATAGACATATTCAGCTTCAA HQMKYQHMEFE'IFCRKLQRTC

TGCCAGCATCCCATAATTGCTTAGTAACCTTCATTCTTTCGGGTAAATAACCAGTCCA ^*PNYHI

GTCCTTACCACCACCAAAAGCCATAACGAACACCTGTGTAGCCGTTGGCTTTATAGT

TGTAGAGGAATTAATTCTTTGTTTGATCAAAGAAAATATTCTTTCAACACCAAATGAAA

TACCAACACATGGAATTTGAGTAGATTTTTTGCCGGAAGCTTCAGAGAACATGTTGA

CCAAATTATCATATC

NMR 104c 249 CATAGGGCAAGGAAGAGCACGTTGACGTTAAAGCAGGACCATTCTCAACCTAGTGTT 250 HRARKSTLTLKQDHSQPSVPSS

CCATCAAGTGTCCATAAAAGCTCGAAAGAGGGGAACATCCTTATTGAGAAAACCACC VHKSSKEGΝILIEKTTDYLVSKP

GATTATCTAGTGTCGAAACCGAAGGCCTCACAGTTATCGAATATGTTCAACAAAAAG KASQLSΝMFΝKKKKRTΝTΝSVD

AAAAAGAGAACAAATACTAATTCTGTAGATGTGTTAGAATATTTTTCATTTGTTTGCGG VLEYFSFVCGDKVPΝYESMGLE

AGATAAAGTACCAAATTATGAATCAATGGGTCTCGAAATATACATTCAAGCTTCCAAA IYIQASKKYKRΝSFTTKVRKSSTI

AAATATAAAAGAAATTCCTTCACTACCAAAGTCCGAAAATCGTCTACCATCTTCGAAG FEVIGFALFLYSTEKKPDΝFEED

TTATTGGATTTGCTTTATTTTTGTATTCTACTGAGAAAAAACCGGACAATTTTGAAGAA GLTVEDISΝPΝΝFSLKIVDEDGE

GATGGATTGACAGTTGAAGACATTTCTAATCCTAATAACTTCTCTTTGAAAATTGTCG PFEDΝFGKLDRKSTIQSISDSEV

ATGAAGACGGTGAACCTTTTGAGGATAACTTTGGTAAACTAGATAGAAAAAGCACTAT VLCKVDDAEKSQΝEIETPLPFET

TCAATCAATTTCGGACAGCGAAGTTGTTTTATGTAAGGTTGATGATGCCGAGAAGTC GGGLMDASTLDAΝSSHDTTDG

TCAAAATGAAATTGAAACACCTTTACCATTTGAAACTGGTGGGGGCCTAATGGATGC TIΝQLSFYKPIIGΝEDDIDKTΝGS

TTCTACTTTGGATGCTAACAGCAGTCACGATACAACTGACGGAACCATCAACCAACT KM DVTVYLYPΝVΝPKFΝ YTTIS V

AAGTTTCTACAAACCTATTATAGGTAATGAAGACGACATCGATAAGACAAACGGTTCG LVTSHIΝDILVKYCKMKΝMDPΝE

AAAATCATTGATGTAACAGTATATTTGTATCCTAACGTGAATCCCAAATTTAACTATAC YALKVLGKΝYILDLΝDTVLRLDGI

TACCATAAGTGTTTTAGTGACCTCACATATAAACGATATTTTGGTAAAATATTGCAAAA ΝKVELISKKDARELHLEKMKPDL

TGAAAAATATGGATCCTAATGAATATGCATTAAAAGTTCTAGGAAAGAACTATATACT KKPVLPTIQSΝDLTPLTLEPLΝS

GGACTTGAATGATACGGTATTAAGGCTCGATGGTATTAATAAGGTGGAACTCATCTC YLKADAGGAVAAIPEΝTKVTSKA

TAAAAAAGATGCAAGGGAGTTGCATTTGGAAAAAATGAAACCTGACCTAAAGAAGCC KKISTKYKLGLAKQHSSSSVASG

CGTTTTGCCAACGATCCAAAGTAATGATCTAACTCCGTTAACTTTGGAGCCTCTGAAC SVSTAGGLAΝGΝGFFKΝ

TCTTATCTCAAAGCAGATGCTGGAGGGGCTGTGGCTGCCATTCCTGAAAACACGAAA

GTTACATCCAAAGCTAAGAAGATATCCACGAAGTATAAGCTGGGCTTGGCAAAGCAG

CATTCGTCATCAAGTGTTGCTAGTGGAAGTGTTTCAACGGCTGGCGGTCTCGCCAAT

GGAAATGGGTTTTTCAAGAAC

1

NDL105W 283 TCGCAATTTCAGACACAACAGAACCGTGACATCAGCACGATGATGGAACACACTAAC 284 SQFQTQQNRDISTMMEHTNSN

AGTAATGATATGAGCGGCTCTGGTAAAAATCTCAAGAAACGTGTATCAAAGGCCTGT DMSGSGKNLKKRVSKACDHCR

GACCATTGCCGAAAAAGAAAAATCAGATGTGATGAAGTGGACCAGCAGACCAAAAAA KRKIRCDEVDQQTKKCSNCIKF

TGTTCAAACTGTATTAAGTTTCAGTTGCCTTGCACTTTCAAACATCGTGATGAGATTC QLPCTFKHRDEILKKKRKLEIKH

TTAAGAAGAAAAGAAAATTAGAAATCAAACATCATGCAACACCAGGGGAATCACTTCA HATPGESLQTSNSISNPVASSS

AACCTCAAATAGTATTAGCAATCCTGTAGCGTCTTCTTCAGTACCGAACAGTGGAAG VPNSGRFELLNGNSPLESNIIDK

GTTTGAACTTTTAAACGGTAATTCCCCCTTAGAAAGCAATATCATCGATAAAGTCTCC VSNIQNNLNKKMNSKIEKLDRK

AATATTCAAAATAATCTTAACAAAAAAATGAATTCAAAGATTGAAAAATTGGATAGAAA MSYIIDSVARLEWLLDKAVKKQE

AATGTCTTACATTATTGACAGTGTGGCTAGACTTGAGTGGTTATTGGACAAGGCTGTT GKYKEKNNLPKPARKIYSTALLT

AAAAAGCAGGAAGGCAAATACAAGGAAAAGAACAATTTGCCCAAACCAGCGAGAAAA AQKLYWFKQSLGVKASNEEFLS

ATATACTCTACGGCACTTTTAACTGCTCAAAAACTCTATTGGTTCAAACAAAGTTTAG PISEILSISLKWYATQMKKFMDL

GAGTGAAAGCGTCCAATGAGGAGTTTCTTTCTCCAATCAGCGAAATATTAAGCATAT SSPAFFSSEIILYSLPPKKQAKRL

CTTTAAAATGGTATGCAACTCAAATGAAAAAATTTATGGATTTGTCATCTCCGGCTTT LENFHATLLSSVTGIISLKECLDL

CTTCTCCAGCGAAATAATATTATACTCATTACCTCCGAAAAAGCAAGCAAAGAGACTT AEKYYSESGEKLTYPEHLLLNV

CTTGAGAATTTTCATGCTACCTTATTATCCTCTGTAACTGGTATAATATCGTTAAAAGA CLCSGASATQSIIRGDSKFLRKD

ATGTCTAGACTTAGCAGAAAAGTACTACAGCGAAAGCGGCGAAAAACTCACATATCC RYDPTSQELKKIENVALLNAMYY

TGAACATTTATTATTAAACGTGTGTCTCTGCTCGGGTGCATCTGCCACTCAATCAATT YHKLSTICSGTRTLQALLLLNRY

ATAAGAGGTGATTCAAAGTTTCTAAGGAAAGATAGATATGATCCAACCTCCCAAGAG FQLTYDTELANCILGTAIRLAVD

TTAAAAAAAATCGAAAACGTTGCCTTACTAAATGCCATGTATTATTATCATAAGCTGTC MELNRKSSYKSLDFEEAIRRRR

CACCATCTGTTCAGGTACAAGAACACTACAAGCTTTATTACTACTGAACCGATATTTT MWWHCF

CAACTTACCTACGATACTGAACTAGCAAATTGTATTTTAGGAACCGCGATTAGATTGG

CGGTTGACATGGAATTAAATAGAAAATCCTCTTACAAGTCACTAGACTTTGAAGAAGC

CATAAGGAGAAGAAGAATGTGGTGGCATTGTTTT

YGR052W 305 AGATTCCTTTCAACTGGAGGATTTTGGCGAGGCGGTACGAATGGCACAATGTCTCG 306 RFLSTGGFWRGGTNGTMSRTI

CACAATCAACAACGTAAATCCTTTCAAATTAAAATTCATACCGAAGACAGTGCCCGCA NNVNPFKLKFIPKTVPAAADSVS

GCTGCAGACTCTGTCTCTCCAGATAGTCAACGTCCAGGTAAGAAGCCCTTCAAATTC PDSQRPGKKPFKFIVSNQSKSS

ATAGTTTCCAATCAGAGCAAGAGTAGCAAGGCTTCTAAAAGCCCGAAGTGGTCGAG KASKSPKWSSYAFPSRETIKSH

CTACGCATTCCCTTCGCGTGAGACCATCAAATCTCATGAGGAGGCCATCAAGAAGCA EEAIKKQNKAIDEQIAAAVSKND

GAATAAAGCTATAGACGAGCAAATAGCTGCTGCAGTATCCAAGAATGACTGCTCTTG CSCTEPPKKRKRKLRPRKALITL

CACAGAACCTCCCAAGAAAAGAAAGAGGAAATTGAGACCAAGAAAGGCGCTGATCA SPKAIKHLRALLAQPEPKLIRVSA

CCCTGAGTCCGAAGGCAATCAAGCATTTAAGGGCACTGCTAGCTCAGCCGGAACCT RNRGCSGLTYDLQYITEPGKFD

AAATTGATTAGAGTTAGCGCTAGAAACCGTGGATGTTCAGGACTAACGTACGATCTA EWEQDGVKIVIDSKALFSIIGSE

CAATATATCACCGAGCCGGGGAAATTCGATGAGGTAGTAGAACAAGATGGCGTTAAA MDWIDDKLASKFVFKNPNSKGT

ATTGTCATCGATTCAAAGGCGTTATTCAGCATCATTGGAAGTGAAATGGACTGGATC CGCGESFMVKPSAPFLEKKNL

GACGACAAGTTGGCCTCTAAGTTTGTCTTCAAGAATCCAAACTCCAAGGGCACATGC PIHLFIHLLIYLHIYHTY^*H^*TLHRG

GGTTGTGGCGAGAGTTTCATGGTTTAAAAACCTTCTGCACCATTTTTAGAAAAAAAGA SVCCS'LFFRCSCRYIFFQNFLE

ATCTACCTATTCACTTATTTATTCATTTACTTATTTATTTACATATTTATCATACATATTA GLLITILNVP^*GAAIFLFNFQQGA

ACATTGAACCCTCCATCGTGGTAGTGTTTGCTGTTCCTAACTTTTCTTTCGTTGTTCT HLKELQIISQ*KAKGRKLMQPAR

TGTAGATATATATTTTTCCAGAATTTTCTAGAAGGGTTATTAATTACAATCTTAAACGT LLYKAL^*SCLNSFIIRSFVTCSFLP

TCCATAAGGGGCCGCGA I I I I I I I GTTCAATTTTCAACAGGGGGCCCATCTCAAAGA LNRISNNKEKRNQLHVP^*NIQNI*

ACTGCAAATTATATCACAGTAAAAGGCAAAGGGGCGCAAACTTATGCAACCTGCCAG TTKRNLQKGL^*RF^*RWLKNWLRI

ATTATTATATAAGGCATTGTAATCTTGCCTCAATTCCTTCATAATTCGTTCCTTTGTCA INIHNYNLYIF*LPSLKRQKMDQS

CTTGTTCCTTTTTACCCTTGAATCGAATCAGCAATAACAAAGAAAAAAGAAATCAACT LTYRI^*LRRAVT

ACACGTACCATAAAATATACAGAATATATGAACGACCAAACGCAATTTACAGAAAGG

GCTCTAACGATTTTGACGTTGGCTCAAAAATTGGCTTCGGATCATCAACATCCACAAT

TACAACCTATACATATTCTAGCTGCCTTCATTGAAACGCCAGAAGATGGATCAGTCC

CTTACCTACAGAATCTAATTGAGAAGGGCCGTTACGA

YHR070W 315 GCGCCAAGACCACATGCTTGTCCTATCTGTCATAGAGCTTTTCACAGACTGGAACAT 316 APRPHACPICHRAFHRLEHQTR

CAGACGAGACACATGAGAATTCATACAGGTGAGAAGCCTCACGCGTGTGACTTCCC HMRIHTGEKPHACDFPGCVKRF

CGGATGTGTGAAAAGGTTCAGTAGAAGCGATGAACTGACGAGACACAGAAGAATTC SRSDELTRHRRIHTNSHPRGKR

ATACAAACTCCCACCCTCGAGGTAAAAGAGGCAGAAAGAAGAAGGTTGTGGGCTCT GRKKKWGSPINSASSSATSIPD

CCAATAAATAGTGCTAGTTCTAGTGCTACCAGTATACCAGATTTAAATACGGCAAATT LNTANFSPPLPQQHLSPLIPIAIA

TTTCACCGCCATTACCACAGCAACACCTATCGCCTTTAATTCCTATTGCTATTGCTCC PKENSSRSSTRKGRKTKFEIGE

GAAAGAAAATTCAAGTCGATCTTCTACAAGAAAAGGTAGAAAAACCAAATTCGAAATC SGGNDPYMVSSPKTMAKIPVSV

GGCGAAAGTGGTGGGAATGACCCATATATGGTTTCTTCTCCCAAAACGATGGCTAAG KPPPSLALNNMNYQTSSASTAL

ATTCCCGTCTCGGTGAAGCCTCCACCTTCTTTAGCACTGAATAATATGAACTACCAAA SSLSNSHSGSRLKLNALSSLQM

CTTCATCCGCTTCCACTGCTTTGTCTTCGTTGAGCAATAGCCATAGTGGCAGTAGAC MTPIASSAPRTVFIDGPEQKQLQ

TGAAACTGAACGCGTTATCGTCCCTACAAATGATGACGCCCATTGCTAGCAGTGCGC QQQNSLSPRYSNTVILPRPRSL

CAAGGACTGTTTTCATAGACGGTCCTGAACAGAAACAACTACAACAACAACAAAATT TDFQGLNNANPNNNGSLRAQT

CTCTTTCACCACGTTATTCCAACACTGTTATATTACCAAGGCCGCGATCTTTAACGGA QSSVQLKRPSSVLSLNDLLVGQ

TTTTCAAGGATTGAACAATGCAAATCCAAACAACAATGGAAGTCTCAGAGCACAAACT RNTNESDSDFTTGGEDEEDGLK

CAGAGTTCCGTACAGTTGAAGAGACCAAGTTCAGTTTTAAGTTTGAACGACTTGTTG DPSNSSIDNLEQDYLQEQSRKK

GTTGGCCAAAGAAATACCAACGAATCTGACTCTGATTTTACTACTGGTGGTGAGGAT SKTSTPTTMLSRSTSGTNLHTL

GAAGAAGACGGACTAAAGGACCCGTCTAACTCTAGTATCGATAACCTTGAGCAAGAC GYVMNQNHLHFSSSSPDFQKE

TATTTGCAAGAGCAATCAAGAAAGAAATCTAAGACTTCCACGCCCACGACAATGCTA LNNRLLNVQQQQQEQHTLLQS

AGTAGATCCACTAGTGGTACGAATTTGCACACTTTGGGGTATGTAATGAACCAAAAT QNTSNQSQNQNQNQMMASSS

CACTTGCATTTCTCCTCATCATCTCCTGATTTCCAAAAGGAGTTGAACAACAGATTAC SLSTTPLLLSPRVNMINTAISTQ

TGAACGTTCAACAACAGCAGCAAGAGCAACATACCCTACTGCAATCACAAAATACGT QTPISQSDSQVQELETLPPIRSL

CAAACCAAAGTCAAAATCAAAATCAAAATCAAATGATGGCTTCCAGTAGTTCGTTAAG PLPFPHMD'YAD

TACAACCCCGTTATTATTGTCACCAAGGGTGAATATGATTAATACTGCTATATCCACC

CAACAAACCCCCATTTCTCAGTCGGATTCACAAGTTCAAGAACTGGAAACATTACCA

CCCATAAGAAGTTTACCGTTGCCCTTCCCACACATGGACTGATACGCTGACAA

YJR090C 367 AGACAAAGGGGGGGTTATCACCATGCGAGACGAAGTAGAGGTGGAGCAGGCGGAT 368 RQRGGYHHARRSRGGAGGSY

CGTATTACAGAGGCGGTAATGCGTCTTATGGCGCGAGATACAACAGTGACTATGAG YRGGNASYGARYNSDYEQPPQ

CAGCCGCCCCAAGAGGGAGACTTGAGACAAACCGGAGCATACTATCGAAACGGATA EGDLRQTGAYYRNGYTDTRPY

CACAGATACAAGGCCGTACTATTCAGCTAATAGCAGGCATTACCAAGCACAACCATC YSANSRHYQAQPSPRYNNGTN

CCCCAGGTACAACAATGGTACAAATTCGTACCACCTACCGCAACGTGGCAATAGTCA SYHLPQRGNSQDTNGRTTSAS

AGATACGAACGGCAGGACCACGAGCGCCTCGCAAGAAGATAATGACGAAAAAAGAG QEDNDEKRVKSRYRNMQADHP

TCAAGTCCAGATACCGGAACATGCAGGCAGATCATCCACGCCAACAACCGATGAGC RQQPMSVGSTSSRNGSSGNSS

GTTGGCAGCACCAGCAGCAGAAATGGTAGTAGTGGGAACAGCAGCACTAGCAGCAC TSSTSNGLPPPPSVSSITNNRSY

CAGCAACGGGCTTCCGCCACCACCTTCAGTATCGTCTATAACCAACAATAGATCATA HSSAYPYSSSHTYNNYHHRETP

TCACAGCAGTGCTTATCCATATTCCAGCAGCCATACTTACAATAATTACCACCACCGT PPPPSNGYYAKGYPVHVPENR

GAGACACCACCGCCTCCCCCATCGAATGGCTATTATGCAAAGGGTTACCCAGTACA SNSDGSSSSWKKKRILDMKDS

CGTACCAGAAAATAGAAGTAACAGCGACGGCTCTAGCAGCAGCGTCGTCAAGAAGA PFIYLTDFDKNVKKTNNTESECE

AAAGAATACTAGATATGAAAGATTCCCCCTTTATTTACTTGACAGATTTTGATAAAAAT KAREVFKESDSIDSALEELNLKI

GTGAAAAAGACAAACAACACCGAAAGTGAATGTGAAAAGGCGAGGGAAGTTTTCAAA NSNELELRLLNNQCDKHALNIQL

GAGAGCGATTCCATCGATTCCGCTTTGGAAGAACTAAACCTAAAAATCAATTCTAATG TQEKLDSLLLMQ*GIPYL*IDKLR

AGCTAGAATTGCGGCTCCTAAATAACCAGTGCGACAAGCATGCATTAAATATTCAAC KRKRRKLKKEIIRK*LAF^*FHESR

TGACCCAAGAAAAGCTGGACTCATTGTTATTAATGCAGTAGGGTATACCGTACCTAT KGTARLSPIR*RARKATRKATTK

AAATAGATAAACTACGTAAACGCAAGCGGCGGAAATTGAAAAAGGAAATTATAAGGA KEKTAKL*KCVSAICLWNRLSICF

AGTGACTAGCGTTTTGATTTCATGAATCGAGAAAAGGTACTGCTAGGTTAAGCCCGA DPLYEERSIV'GSSSARHVLPRN

TTAGGTAAAGAGCCAGGAAAGCGACTAGGAAAGCGACTACGAAAAAAGAAAAAACT PTHTTHGLFRLPPRIITIPEQTHT

GCGAAACTGTGAAAGTGCGTGTCAGCCATCTGTCTTTGGAATCGATTATCTATTTGC FEFASRRV

TTTGATCCCCTTTATGAAGAACGCAGCATTGTGTGAGGCTCTTCCTCTGCTCGCCAC

GTGCTCCCACGAAATCCCACCCACACCACACACGGTCTGTTTCGTCTTCCCCCCCG

CATTATTACTATCCCCGAGCAAACTCACACTTTTGAATTCGCGTCGCGTCGCGTCG

YNL260C 409 AGATTTCCTCCGGATAATGACCAAAGACCCTTTAGATGTGAAATTTGTTCACGAGGTT 410 RFPPDNDQRPFRCEICSRGFHR

TCCACAGACTTGAACATAAAAAAAGGCACGGAAGAACGCACACTGGCGAGAAGCCT LEHKKRHGRTHTGEKPHKCTV

CACAAATGTACCGTTCAGGGCTGTCCGAAAAGCTTCAGCCGAAGCGATGAACTAAAA QGCPKSFSRSDELKRHLRTHTK

AGACATTTGAGGACACATACTAAAGGCGTCCAAAGGCGCAGAATAAAATCCAAGGG GVQRRRIKSKGSRKTWNTATA

CTCGCGAAAAACCGTTGTGAATACTGCTACCGCCGCCCCTACCACCTTCAATGAAAA APTTFNENTGVSLTGIGQSKVP

CACTGGTGTTTCGCTCACGGGGATAGGTCAATCTAAAGTGCCACCTATTCTTATCTC PILISVAQNCDDVNIRNTGNNNG

CGTTGCTCAGAATTGCGATGACGTGAATATACGAAATACTGGAAATAATAATGGCATT IVETQAPAILVPVINIPNDPHPIPS

GTGGAGACACAGGCACCTGCAATTTTAGTGCCTGTGATAAATATTCCAAATGACCCT SLSTTSITSIASVYPSTSPFQYLK

CATCCGATTCCAAGTAGCCTCTCCACTACTTCTATCACCTCCATTGCATCAGTATATC SGFPEDPASTPYVHSSGSSLAL

CCTCTACTTCTCCATTCCAGTACCTGAAAAGCGGGTTTCCTGAAGATCCTGCATCTA GELSSNSSIFSKSRRNLAAMSG

CACCGTATGTACATTCGTCCGGAAGTTCTTTAGCCCTGGGTGAATTGTCTTCAAACT PDSLSSSKNQSSASLLSQTSHP

CCTCTATATTTTCGAAATCTAGGAGGAATTTGGCCGCCATGAGTGGTCCTGATTCTTT SKSFSRPPTDLSPLRRIMPSVNT

GAGTAGTTCTAAAAACCAATCCAGTGCTTCGCTTCTTTCTCAAACTTCACATCCATCA GDMEISRTVSVSSSSSSLTSVTY

AAGAGCTTTTCAAGACCGCCAACAGACTTAAGTCCTCTGCGAAGAATCATGCCTTCT DDTAAKDMGMGIFFDRPPVTQK

GTAAACACAGGAGACATGGAAATTTCAAGGACAGTATCCGTTTCGAGCAGTTCATCA ACRSNHKYKVNAVSRGRQHER

TCACTCACTTCTGTTACGTATGATGACACCGCGGCTAAAGACATGGGCATGGGAATA AQFHISGDDEDSNVHRQESRAS

TTTTTTGATAGGCCACCTGTAACACAGAAAGCTTGCAGGAGCAATCATAAGTACAAG NTSPNVSLPPIKSILRQIDNFNSA

GTTAATGCTGTTAGCAGAGGGAGACAACATGAAAGGGCACAATTTCATATATCTGGA PSYFSK^*TSSK^*NV^*NIRRKITMG

GATGATGAGGACAGTAACGTTCACCGCCAAGAATCAAGAGCATCCAACACAAGTCC S*NLIYFFLE^*RRKENLWRKSLQI

CAATGTATCATTGCCTCCGATAAAGAGCATTTTGCGACAAATTGATAATTTCAACAGT *KLCFKKPM*D*NMVQWSSSGP

GCTCCTTCTTACTTCAGTAAATAAACAAGCAGTAAATAAAACGTATAGAATATAAGAA VQSPALPSIICLYR'NTGHAIFFV

GGAAAATAACAATGGGTTCTTGAAACTTGATATACTTTTTCCTGGAATGAAGACGCAA ALIFILKRPICSPLEKLTKKRGKE

AGAAAACCTTTGGAGAAAATCATTGCAAATTTAAAAGCTGTGCTTCAAAAAACCCATG NNTDLVGFGPLL^*SQVESILDKS

TGAGACTGAAACATGGTCCAGTGGTCTTCATCCGGACCGGTTCAAAGTCCTGCTCTA YYILYKWSCTIINYLH^*NMAGDN

YMR065W 455 TTATCAGATGAGGAAGAACGAATGAGAGAAAAAATGTCCATACGCAAAGCTAGTGCT 456 LSDEEERMREKMSIRKASALEW

CTCGAGTGGGAAAGATTTTTGCTTACCGACTTCAGTGCGATAATTGACCTTTTCCAA ERFLLTDFSAIIDLFQGQYASRL

GGACAGTACGCATCTAGGCTACAATGTCAAGTTTGTGAACATACCTCCACAACTTAC QCQVCEHTSTTYQTFSVLSVPV

CAAACATTCTCTGTTCTTTCTGTTCCTGTCCCACGCGTTAAAACTTGTAACATATTAG PRVKTCNILDCFREFTKCERLGV

ACTGTTTCCGGGAATTCACCAAATGCGAAAGGTTAGGTGTCGATGAACAATGGTCAT DEQWSCPKCLKKQPSTKQLKIT

GTCCTAAATGCTTAAAAAAGCAGCCTTCCACTAAACAACTGAAGATTACTAGATTGCC RLPKKLIINLKRFDNQMNKNNVF

TAAGAAACTAATTATTAATTTGAAACGATTTGACAATCAAATGAATAAGAATAATGTGT VQYPYSLDLTPYWARDFNHEAI

TTGTCCAATATCCTTACTCCTTAGATCTTACACCATATTGGGCGAGAGATTTTAATCA VNEDIPTRGQVPPFRYRLYGVA

TGAAGCTATTGTTAATGAGGACATTCCTACCAGGGGCCAAGTACCACCATTTAGATA CHSGSLYGGHYTSYVYKGPKK

CAGACTGTATGGGGTTGCATGTCATTCGGGGAGTTTGTATGGGGGACACTATACTTC GWYFFDDSLYRPITFSTEFITPS

ATACGTTTATAAGGGACCAAAAA GGTTGGTATTTTTTCGATGACTCGCTTTATCGT AWLFYERIF*IKLPVCNT^*MAHIE

CCTATAACGTTTAGTACTGAATTTATCACACCCAGTGCATATGTTTTATTTTACGAAAG AQIQSAYYILNRSTIIYNTNNN^*R

AATTTTTTGAATCAAATTACCGGTTTGCAACACATGAATGGCGCACATTGAGGCACAA NNESMYILILIYLCYGLHT^*RPITI

ATACAATCTGCATATTATATACTTAACAGAAGTACAATCATATACAATACAAACAACAA SWAHWKRRDAANSAFPLVCLR

TTGACGAAACAATGAAAGTATGTACATCCTTATACTTATCTATTTGTGTTATGGGCTA AFTTASLGKPRSISCLIVCSGNV

CATACGTAGAGGCCGATCACAATATCAGTGGTTGCTCATTGGAAAAGGAGGGATGC LPVAWAPAVLLPVGVGPIPVLG

AGCAAATTCTGCATTTCCGTTAGTCTGTTTAAGAGCCTTTACCACAGCATCCCTAGG ASAGTAPGCGGRVELVLVGVR

GAAACCTAGATCCATAAGCTGTTTAATCGTTTGTTCGGGAAACGTTCTCCCGGTTGC GLSDEWTDVGAGSPS*KLFGIS

CGTTGTAGCACCGGCTGTACTTCTTCCCGTAGGTGTTGGCCCCATACCAGTCCTTG ASLKKLVSTSAILRTFSFKSTQA

GAGCAAGCGCAGGAACAGCCCCCGGTTGTGGTGGTAGAGTTGAACTAGTCTTGGTG KCLFSMSSPIKTSMSV

GGTGTTAGGGGCTTATCAGACGAAGTTGTAACTGACGTTGGTGCTGGAAGTCCTTCT

TGAAAACTTTTTGGGATTTCCGCTTCACTCAAAAAGCTTGTCTCCACTTCTGCTATCC

TAAGAACGTTTTCCTTTAAGTCCACACAAGCCAAATGCCTTTTCAGCATGTCTAGTCC

TATCAAAACATCGATGTCAGTA

_6l

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YMR065W 479 GTGGAAACGAAAGAGCATGAACTGACACAGGCTGAATTAAACCTAAAGGATGCTATT 480 VETKEHELTQAELNLKDAIGFDP

GGGTTCGACCCCACATGGATTGGAAATATGCTAGCCACGGTCGAGTTATACTACCA TWIGNMLATVELYYQRGHYDKA

GCGCGGTCATTATGACAAGGCTTTGGAAACATCCGACCTTTTTGTAAAGAGTATTCA LETSDLFVKSIHAEDHRSGRQS

TGCTGAAGACCACAGGTCTGGTAGGCAATCCAAACCCAATTGCTTGTTTTTACTTTTA KPNCLFLLLRAKLLYQKENYMA

AGAGCGAAATTGCTTTATCAGAAGGAAAATTATATGGCCAGTCTGAAAATTTTTCAAG SLKIFQELLVINPVLQPDPRIGIG

AATTGCTGGTCATCAACCCTGTTTTGCAACCAGATCCTCGTATTGGAATTGGTCTGT LCFWQLKDSKMAIKSWQRALQL

GTTTTTGGCAATTAAAGGATTCTAAGATGGCTATCAAATCGTGGCAGAGAGCCTTAC NPKNTSASILVLLGEFRESFTNS

AACTAAATCCAAAAAATACGAGTGCCTCCATTTTAGTATTATTGGGTGAATTTCGCGA TNDKTFKEAFTKALSDLNNIFSE

ATCTTTCACCAATTCTACAAACGATAAAACATTCAAGGAGGCTTTTACAAAGGCATTG NQHNPVLLTLLQTYYYFKGDYQ

AGTGACTTGAACAACATTTTCTCCGAAAACCAGCATAATCCTGTTTTATTGACGTTAT TVLDIYHHRILKMSPMIAKIVLSE

TACAAACGTACTACTACTTCAAAGGTGATTATCAAACTGTTTTGGATATTTATCACCAT SSFWCGRAHYALGDYRKSFIMF

AGGATTTTAAAAATGAGTCCGATGATAGCCAAAATTGTTCTATCTGAATCTTCATTTTG QESLKKNEDNLLAKLGLGQTQIK

GTGTGGTAGAGCGCATTATGCATTAGGTGATTATCGTAAATCATTCATCATGTTTCAA NNLLEESIITFENLYKTNESLQEL

GAAAGTTTAAAGAAAAATGAAGATAACCTGCTCGCGAAATTAGGGCTGGGTCAAACC NYILGMLYAGKAFDAKTAKNTS

CAAATTAAAAATAACTTGTTGGAGGAAAGTATCATCACTTTCGAGAATCTTTACAAGA AKEQSNLNEKALKYLERYLKLTL

CAAATGAAAGTTTACAAGAATTAAACTATATCTTGGGTATGCTTTATGCAGGCAAGGC ATKNQLVISRAYLVISQLYELQN

CTTTGATGCCAAGACAGCCAAAAACACTTCAGCAAAAGAACAAAGCAACTTAAATGA QYKTSLDYLSKALEEMEFIKKEI

AAAAGCTCTAAAGTATTTGGAGAGATACCTAAAATTAACGCTTGCCACAAAGAACCAA PLEVLNNLACYHFINGDFIKADD

TTAGTTATATCTAGAGCTTACCTAGTTATCTCTCAGCTGTATGAATTACAAAACCAATA LFKQAKAKVSDKDESVNITLEYN

TAAAACTTCATTGGATTATCTTTCCAAAGCTTTGGAGGAAATGGAATTCATTAAAAAG IARTNEKNDCEKSESIYSQVTSL

GAAATTCCGTTGGAAGTGCTCAATAATTTGGCATGTTACCACTTCATTAACGGTGACT HPAYIAARIRNLYLKFAQS

TTATAAAGGCTGATGATCTCTTTAAGCAAGCAAAGGCAAAAGTGAGTGATAAAGATG

AAAGTGTCAATATTACACTTGAATATAACATAGCAAGAACTAATGAGAAAAACGACTG

TGAAAAATCAGAATCTATATATTCCCAGGTGACGTCTTTACATCCAGCTTACATTGCG

GCAAGAATAAGAAATCTATATCTCAAGTTTGCACAATCTAA

YOL033W 507 ACGGACCCTTTGGATTCCGTGGTCGATCAAGTGGTCAACCTCAACTTTCACACGTAC 508 TDPLDSWDQWNLNFHTYCLT

TGTTTGACAGAGCACATACCAAGAATTGAGGCCAAGTTTATATACCCCGAAGAGCAG EHIPRIEAKFIYPEEQSLGKNPEE

TCATTGGGCAAGAATCCTGAGGAAGTCATAACCAAGCTAGAAACATCGTTCAAGAAT VITKLETSFKNFMSHAQEIKTRY

TTCATGAGTCATGCGCAAGAAATCAAGACTCGTTATGCTGATAGACCCGATGTGCGG ADRPDVRTKFIIGMEIESCDMAH

ACTAAATTCATTATAGGAATGGAGATCGAAAGTTGTGACATGGCTCATATCGAATATG IEYAKRLMKENNDILKFCVGSVH

CAAAGCGACTCATGAAGGAGAATAATGATATTTTGAAGTTTTGTGTGGGTTCGGTCC HVNGIPIDFDQQQWYNSLHSFN

ATCACGTCAACGGGATCCCTATTGATTTCGACCAACAACAATGGTACAATTCATTGCA DNLKHFLLSYFQSQYEMLINIKP

TTCCTTCAATGATAATTTGAAACATTTTCTCCTGTCTTACTTCCAATCACAGTACGAAA LWGHFDLYKLFLPNDMLVNQK

TGCTGATCAATATTAAACCGTTGGTCGTGGGTCACTTCGACCTTTACAAATTATTTTT SGNCNEETGVPVASLDVISEWP

GCCCAATGACATGCTAGTAAACCAGAAATCGGGCAACTGCAACGAAGAAACCGGAG EIYDAWRNLQFIDSYGGAIEINT

TTCCTGTAGCTTCACTGGACGTCATCAGTGAATGGCCAGAAATATACGATGCAGTTG SALRKRLEEPYPSKTLCNLVKK

TAAGAAATTTACAATTTATAGACTCCTATGGCGGCGCAATTGAAATCAATACGTCCGC HCGSRFVLSDDAHGVAQVGVC

ATTAAGAAAGCGCCTCGAGGAGCCGTACCCCAGCAAAACCTTATGTAATCTGGTCAA YDKVKKYIVDVLQLEYICYLEES

GAAGCACTGTGGATCCAGATTTGTTCTAAGTGATGACGCACACGGCGTGGCGCAAG QSPENLLTVKRLPISQFVNDPF

TGGGTGTGTGCTATGACAAGGTAAAGAAATACATAGTAGACGTGCTACAATTAGAGT WAN l*SDYLELDVHKKKLSYKLM

ATATTTGCTACCTTGAGGAAAGCCAATCACCAGAGAATCTGTTAACTGTAAAGAGATT YSCVEVNDLSKKKISH^*ID*IDCS

ACCCATTTCGCAATTCGTTAATGATCCCTTTTGGGCCAATATATAATCTGATTATCTTG FYIGCPNCLSLLFV"QHPVCSPT

AACTGGATGTACATAAAAAAAAACTGTCATATAAACTCATGTACAGCTGTGTAGAAGT RPHLEVQHERV^*TIEEI^*NGFGS

G TGATCTTTCC GAAAAAAATTTCACACTGAATTGATTGAATTGACTGCTCATTTT RVAKKSKTKAVSHAMGINNPIPR

ATATTGGGTGCCCAAATTGTCTTTCTTTATTGTTCGTTTAGTAGCAGCACCCTGTTTG SLKSETKWLYGFGWETFKKVL

CTCGCCGACTCGGCCGCATCTGGAAGTTCAACATGAAAGAGTATAAACTATCGAAGA P*FVHHLLTSNKQLIKKSSEDFG^*

AATCTAGAACGGCTTTGGATCACGGGTGGCAAAGAAATCAAAAACAAAAGCTGTATC FCQTKPSFWCGPSHSS

ACACGCAATGGGTATTAACAATCCTATTCCAAGGAGTTTAAAGAGTGAGACAAAGTA

TGTTTTATACGGGTTTGGCTGGGAAACCTTTAAAAAAGTTCTCCCTTAATTTGTTCAT

ill

© © α. ω

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VIZ

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YOL033W 535 GCAAACGACATTGTGACCCTGGCAAATTTGCAATATAATGGCAGTACACCTGCAGAT 536 ANDIVTLANLQYNGSTPADAFET

GCATTTGAAACAAAAGTCACAAACATTATCGACAGACTGAACAATAATGGCATTCATA KVTNIIDRLNNNGIHINNKVACQL

TCAATAACAAGGTCGCATGCCAATTAATTATGAGAGGTCTATCTGGCGAATATAAATT IMRGLSGEYKFLRYTRHRHLNM

TTTACGCTACACACGTCATCGACATCTAAATATGACAGTCGCTGAACTGTTCTTAGAT TVAELFLDIHAIYEEQQGSRNSK

ATCCATGCTATTTATGAAGAACAACAGGGATCGAGAAACAGTAAACCTAATTACAGG PNYRRNPSDEKNDSRSYTNTTK

AGAAATCCGAGTGATGAGAAGAATGATTCTCGCAGCTATACGAATACAACCAAACCC PKVIARNPQKTNNSKSKTARAH

AAAGTTATAGCTCGGAATCCTCAAAAAACAAATAATTCGAAATCGAAAACAGCCAGG NVSTSNNSPSTDNDSISKSTTEP

GCTCACAATGTATCCACATCTAATAACTCTCCCAGCACGGACAACGATTCCATCAGT IQLNNKHDLHLRPETY^*IYSKSY^*

AAATCAACTACTGAACCGATTCAATTGAACAATAAGCACGACCTTCATCTTAGGCCAG SF^***TPWTPPSRFRSITNPYKIC

AAACTTACTGAATCTACAGTAAATCATACTAATCATTCTGATGATGAACTCCCTGGAC SSHTLSII*SΗKRS^*CSKKKYTN^*

ACCTCCTTCTCGATTCAGGAGCATCACGAACCCTTATAAGATCTGCTCATCACATACA RYW^*PTISLPGQHQNINKGIAHS

CTCAGCATCATCTAATCCTGACATAAACGTAGTTGATGCTCAAAAAAGAAATATACCA *HSL*LTQFE^*IGCSRYHSMLYQK

ATTAACGCTATTGGTGACCTACAATTTCACTTCCAGGACAACACCAAAACATCAATAA RLRTV*RHCTCTYRKIWRLLLGI*

AGGTATTGCACACTCCTAACATAGCCTATGACTTACTCAGTTTGAATGAATTGGCTGC KVLASIKYLRTHHQ^*CPYK*KYT

AGTAGATATCACAGCATGCTTTACCAAAAACGTCTTAGAACGGTCTGACGGCACTGT QISLSFHSSNACACQCTDNSILT

ACTTGCACCTATCGTAAAATATGGAGACTTTTACTGGGTATCTAAAAAGTACTTGCTT *KΗHHVF^*RIRCRLV*CY^*LSMS*

CCATCAAATATCTCCGTACCCACCATCAATAATGTCCATACAAGTGAAAGTACACGCA LFNRQKHQTQTYQRFTTKIPKFI

AATATCCTTATCCTTTCATTCATCGAATGCTTGCGCATGCCAATGCACAGACAATTCG RTLSIPTY*HIWSSSQPTKKCTIL

ATACTCACTTAAAAATAACACCATCACGTATTTTAACGAATCAGATGTCGACTGGTCT FHLIY"DNKIPLGLSITRPSR

AGTGCTATTGACTATCAATGTCCTGATTGTTTAATCGGCAAAAGCACCAAACACAGAC

ATATCAAAGGTTCACGACTAAAATACCAAAATTCATACGAACCCTTTCAATACCTACA

TACTGACATATTTGGTCCAGTTCACAACCTACCAAAAAGTGCACCATCCTATTTCATC

TCATTTACTGATGAGACAACAAAATTCCGTTGGGTTTATCCATTACACGACCGTCGC

GA

YER057C 543 TATGGAGTTAATTTTTTCAACAACACAGGGGTGGACAACTCTTACTTGGTTTCTTTTA 544 YGVNFFNNTGVDNSYLVSFISY

TCAGCTATGCCGTCAACGTCGCCTTCAGTATACCGGGTATGTATTTAGTGGATCGAA AVNVAFSIPGMYLVDRIGRRPVL

TTGGTAGAAGACCAGTCCTTCTTGCTGGAGGTGTCATAATGGCAATAGCAAATTTAG LAGGVIMAIANLVIAIVGVSEGKT

TCATTGCCATCGTTGGTGTTTCCGAGGGAAAAACTGTTGTTGCTAGTAAAATTATGAT WASKIMIAFICLFIAAFSATWGG

TGCTTTTATATGCCTTTTCATTGCTGCATTTTCGGCGACATGGGGTGGTGTCGTGTG WWWSAELYPLGVRSKCTAIC

GGTGGTATCTGCTGAACTGTACCCACTTGGTGTCAGATCGAAATGTACCGCCATATG AAANWLVNFTCALITPYIVDVGS

CGCTGCCGCAAATTGGCTAGTTAATTTCACCTGTGCCCTGATTACACCTTACATTGTT HTSSMGPKIFFIWGGLNWAVIV

GATGTCGGATCACACACTTCTTCAATGGGGCCCAAAATATTCTTCATTTGGGGCGGC VYFAVYETRGLTLEEIDELFRKA

TTAAATGTCGTGGCCGTTATCGTTGTTTATTTCGCTGTTTATGAAACGAGGGGATTGA PNSVISSKWNKKIRKRCLAFPIS

CTTTGGAAGAGATTGACGAGTTATTTAGAAAGGCCCCAAATAGCGTCATTTCTAGCA QQIEMKTNIKNAGKLDNNNSPIV

AATGGAACAAAAAAATAAGGAAAAGGTGCTTAGCCTTTCCCATTTCACAACAAATAGA QDDSHNIIDVDGFLENQIQSNDH

GATGAAAACTAATATCAAGAACGCTGGAAAGTTGGACAACAACAACAGTCCAATTGT MIAADKGSGSLVNIIDTAPLTSTE

ACAGGATGACAGCCACAACATAATCGATGTGGATGGATTCTTGGAGAACCAAATACA FKPVEHPPVNWDLGNGLGLNT

GTCCAATGATCATATGATTGCGGCGGATAAAGGAAGTGGCTCGTTAGTAAACATCAT YNRGPPSIISDSTDEFYEENDSS

CGATACTGCCCCCCTAACATCTACAGAGTTTAAACCCGTGGAACATCCGCCAGTAAA YYNNNTERNGANSVNTYMAQLI

TTACGTCGACTTGGGGAATGGTTTGGGTCTGAATACATACAATAGAGGTCCTCCTTC NSSSTTSNDTSFSPSHNSNART

TATCATTTCTGACTCTACTGATGAGTTCTATGAGGAAAATGACTCTTCTTATTACAATA SSNWTSDLASKHSQYTSPQ*KP

ACAACACTGAACGAAATGGAGCTAACAGCGTCAATACATATATGGCTCAACTAATCA IASYDRSRSYKPFYINYSYTRTL

ATAGCTCATCTACTACAAGCAACGACACATCGTTCTCTCCATCACACAATAGCAATGC EFLLTFQHDRHRKEL*ITHNITHK

AAGAACGTCCTCTAATTGGACGAGTGACCTCGCTAGTAAGCACAGCCAATACACTTC *PMSLIFPR*FGVKWNTKWVN^*S

CCCCCAATAAAAACCAATAGCATCTTACGATCGTTCGAGGTCTTATAAACCGTTTTAT ^*LLPG*STSCGTLTMYGFLNHRV

ATAAATTATTCTTATACACGCACTCTTGAGTTTCTTTTAACATTCCAACATGATAGACA PDAGCCCS

TAGAAAAGAATTATAGATAACACATAACATTACTCACAAGTAGCCCATGAGTTTGATT

TTTCCTCGTTAATTCGGTGTCAAGTGGAATACAAAATGGGTCAACTGATCATAACTAT

mz cs

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YOR057W 617 AGCTTCAAGTTAAAAACTTCACCTATACGGAAACATTTTCATGTTAAACCCAAAAGAA 618 SFKLKTSPIRKHFHVKPKRITRV

TAACAAGGGTGAGGACAGGAAGGGTATCACACAATATAATTGAAAAGAAATACCGTT RTGRVSHNIIEKKYRSNINDKIEQ

CCAATATTAATGATAAAATTGAACAATTAAGGAGAACTGTCCCAACGCTACGAGTAGC LRRTVPTLRVAYKKCNDLPITSR

GTATAAAAAATGTAACGATCTTCCCATTACATCTAGAGACCTAGCAGACTTAGATGGC DLADLDGLEPATKLNKASILTKSI

CTAGAGCCAGCAACAAAATTGAATAAGGCGTCCATTTTAACGAAATCTATTGAATATA EYICHLERKCLQLSLANQHLSND

TTTGTCATTTAGAAAGAAAGTGCCTACAGCTAAGCTTGGCTAATCAACATTTATCAAA TRDSFVHLTEPSQPLSDNSSSE

TGACACACGGGATTCTTTCGTCCATCTTACCGAGCCATCTCAACCGTTGAGTGACAA QVQKQTRSCQRQRQRQPRQQ

CAGTAGTAGTGAACAAGTACAAAAGCAAACCAGAAGCTGCCAACGCCAACGGCAGC QPLHNIQYNIPHQNGLMSGTNN

GGCAGCCACGACAACAACAACCATTGCATAACATTCAATATAATATTCCACATCAAAA SHDMDFNNAGDF^*YLNNKMFQ

CGGCCTCATGAGCGGCACCAACAATTCACATGATATGGATTTTAACAACGCAGGAGA HDR*NEEEILYTNLYLCVCVTL*F

CTTTTAATACTTGAACAACAAAATGTTTCAACACGACAGATAAAACGAAGAAGAAATA *FCIEKSILFISFSVSKHSQMP*AP

CTATATACAAATCTGTACTTGTGTGTATGCGTAACATTGTAATTTTAATTTTGCATTGA HRNLV*FISLSAV^*IITI*KKDSGKK

GAAATCTATTTTATTCATCTCTTTTAGTGTTAGTAAACACTCGCAGATGCCTTGAGCT KLYLIFSSCS*INWN*G*NMMMY

CCGCACCGTAATTTAGTTTAATTTATTTCTCTTAGTGCCGTCTAAATAATAACGATCTA L"VKSRGHLTFFTIAFTLFTSAR

AAAAAAAGATTCCGGGAAAAAGAAACTATACCTTATATTTAGTTCATGCAGTTAAATA GFAFAFFTTTLTLRASGAVITFAL

AATGTTGTTAATTAAGGGTAAAATATGATGATGTATTTATGATAAGTAAAGAGTAGGG FTARTAWKAGAPTMTRVLTTFA

GTCACCTAACATTTTTCACAATTGCATTTACTCTTTTCACAAGTGCACGTGGTTTCGC LTVTYFYSHIY*VLFDIWFSVISL

ATTTGCAI I I I I CACCACAACTCTTACATTGAGGGCTTCCGGTGCTGTGATCACATTT RYS^*GGRCAPFLYRPRLITTSQP

GCACTTTTCACCGCCAGAACAGCTTGGAAGGCAGGTGCTCCCACAATGACAAGAGT SRIPILFLSERQKSLPHSNKDAFI

CCTTACAACATTCGCCTTAACAGTCACATA I I I I I ACAGTCATATTTATTGAGTACTAT VLFLKSACSRGFSKRKKYMTGC

TTGACATTGTAGTGTTCAGTGTAATATCGCTTCGCTATTCTTAGGGGGGACGATGTG YAΓRKVELILKGFCTEVNARRIW

CGCCCTTCTTATATAGGCCCCGCTTGATCACTACTTCACAGCCCTCTCGCATCCCTA NTΎRTERRTKIMLVEYGITVN^*S

TCC I I I I I I I GTCCGAAAGACAAAAATCACTTCCACACAGCAATAAAGACGCATTTAT FIRDVDDGI*FSCISDLKIKAINDH

TGTTCTTTTTCTTAAAAGTGCTTGCAGTAGGGGGTTTAGCAAAAGAAAAAAATATATG KK'RQLYFCSYVFSSTGPALITIF

YOR057W 621 TGGCCAAACAAGAATGAAAAAAACCACACAGTTAAAAGAGCGTTATCAACGGATATG 622 WPNKNEKNHTNKRALSTDMTS

ACCAGCAATATTTTGAGTAGCACAAACGCGAGCTCAAACGAAGAAAATTCTAGGAGT ΝILSSTΝASSΝEEΝSRSSSAAΝ

TCTTCTGCAGCCAATGTACGCTCCGGAACAGGTGCAAATACACTTACTAATGGCGGC VRSGTGAΝTLTΝGGSTRKRLAC

AGTACTAGAAAGAGACTTGCGTGCACTAATTGTAGAAATAGAAGGAAAAAATGTGAT TΝCRΝRRKKCDLGFPCGΝCSR

TTAGGATTTCCCTGTGGTAACTGTTCCAGGTTGGAATTGGTTTGTAATGTTAATGACG LELVCΝVΝDEDLRKKRYTΝKYV

AGGACTTAAGAAAGAAGCGGTACACTAATAAATATGTCAAGTCTTTGGAGAGCCATA KSLESHIAQLETΝLKΝLVQKIYP

TTGCTCAACTGGAGACCAACTTAAAAAACCTAGTTCAGAAGATCTACCCTGACGATG DDEQILΝRMMVGDVLSALPDSS

AGCAAATACTGAACCGAATGATGGTAGGTGATGTATTATCAGCTCTACCAGACAGTT QVSIΝYTDQTPSLPIPATRGTFII

CACAAGTCTCAATCAATTATACTGACCAAACTCCCTCTCTTCCAATTCCCGCAACCAG EΝDKVSQPLSSFΝQQTEPSTLΝ

AGGTACATTCATTATTGAAAACGATAAGGTCAGTCAACCTCTATCGTCCTTTAACCAA SGIFΝTQKQΝFEESLDDQLLLR

CAAACAGAGCCATCTACTCTAAACTCGGGTATCTTCAACACCCAAAAACAAAATTTCG RSLTPQGEKKKKPLVKGSLYPE

AAGAATCCCTTGATGATCAGTTACTΠTACGAAGATCGTTAACACCGCAAGGTGAAAA GPVSYKRKHPVKSDSLLPVSSL

AAAGAAGAAACCGTTGGTAAAAGGTAGTCTTTATCCTGAAGGACCTGTCAGTTACAA TAATDPSTFSDGITAGΝSVLVΝ

ACGGAAGCACCCCGTAAAATCGGACAGTTTATTGCCTGTGTCTTCGTTAACAGCTGC GELKKRISDLKTTNIVRGLΝDDΝ

TACAGACCCATCTACTTTTTCTGACGGTATAACTGCTGGTAATTCCGTCCTAGTTAAT PΝSIΝΝDPRILKSLSΝFYKWLYP

GGTGAACTGAAAAAACGTATATCCGACTTGAAGACCACCGTAATAGTAAGAGGACTA GYFIFVHRESFLYGFFΝHSKΝΝ

AACGATGATAATCCCAACTCTATCAATAACGATCCCAGGATTTTAAAATCTCTTTCCA YEDSSYCSVELIYAMCAVGSRL

ATTTCTATAAGTGGCTGTATCCAGGTTATTTTAI I I I I GTTCACAGAGAAAGTTTTCTT TPDLQEYSEVYYQRSKKTLLQL

TATGGATTCTTCAATCACTCCAAGAATAACTATGAAGACTCAAGTTACTGTTCTGTCG VFDEQSTARITTNQALFCLAFYE

AATTGATATATGCCATGTGCGCTGTCGGTTCGAGGCTTACACCGGATCTACAAGAAT LGKGΝΝQLGWYFSGLAIRVGYD

ATTCAGAAGTGTATTATCAACGAAGTAAGAAGACTTTATTACAGCTCGTCTTTGATGA MGFQLDPKVWYNDDΝΝLQLTQ

GCAAAGTACAGCCCGTATCACCACCGTCCAAGCGCTATTTTGCTTAGCCTTTTACGA SELEIRSRIYWGCYIADHFICLML

ACTGGGGAAGGGTAATAATCAATTAGGGTGGTACTTCTCGGGCCTGGCTATCAGGG GRTSTLSVSΝSTMPESDELPEV

TCGGCTACGACATGGGATTTCAACTGGACCCTAAAGTCTGGTACGTTGATGATAACA ΝGTEEFRFIGRHVLQISLPLKΝLI

693 s

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H >INHllS>13NN>iy3>ISN>πλM9 U 910V90VWW9V91W10W9W09V1V0W1V1W9110V0VJJ.V1W01919VW α. λdsyi3ταs iNταττ>iNiΛj3isiΛiα 91V9WV1V919V919WW099WV09W09WJJJ.0101V91V91099VW1V11 sα3i»sDVd>nΛTNaιawiΛVdH 10V91990W11V0110V00V1191V099111W1W010919W991190011VW9

TSlΛDSΛIS3S»30αN l3NiyVI 9VVWVJJ.V0JJ.VV991VVV99V99JJJ.09VVV00JJJ.01VJJ.V99JJ.V0JJ.0VVVV1

Nλ3TllNΛS3α>iαSΛ >ιNOXdT V1W00WW0V11W91V19109V010101V119V100V1109V9V101V1V119V11 ααV»ldα9NldHΛ0VTNNTΛ3Td W00W9WV0V00911090W11WW100V1V9V9V99111V19VW10109WW

V91VW110W09WV0W9VWV09V0110V0WVW009V0V9W0091V91110

NDT3ΛlQSIΛTΛVaSIΛlDN llV 099W099V091VJJJ.091V19991101V1V10WV11W9W0V1119VW91VW0

TlT»TAd3TA>nV>l3NTNSD3 IV V9W9V.IJJ.01W9V99JJJ.0V01V01V19VW99V99119119W1WWV.IJ.WV0

SlNWl αdV>l9VλTVM91lλN 00VW0199910999V11WV9090109100W1V9W91WWV9VW1119VW9

T3DTS3Nl>IΛTN3dlllS33TTNN W011191V01V011V01VW1901V11V9199V11V091V11V0909V9V1991919

»IDlQ9T9T>IVTTNα3N>l>nS3D 91111V01101W9101V1011911WW009V1V91V90019V91WWV I I I I V99V dlΛlldS l dΛα9TVλHV d90MdSS 1V00V01V111V1V991111910VW01V11V9199VW0110V10V10V190VW0V1

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YGL108C 695 GCTGATGCGTTTGGGCCCAAAGCCCAAAGAAAGAGACCACGTCTTGCTGCATCCAA 696 ADAFGPKAQRKRPRLAASNLED

TCTAGAGGACTTGGTCAAGGCTACAAATGAAGACATTACCAAGTATGAGGAAAAGCA LVKATNEDITKYEEKQVLDATLG

AGTCTTAGATGCCACATTAGGACTAATGGGGAACCAGGAAGACAAAGAAAATGGGT LMGNQEDKENGWTSAAKEAIFS

GGACCTCCGCAGCAAAAGAAGCTATTTTCAGTAAGGGTCAATCCAAACGTATTTGGA KGQSKRIWNELYKVIDSSDWIH

ACGAATTATATAAAGTCATCGATTCGTCTGACGTGGTAATACATGTTCTAGATGCAAG VLDARDPLGTRCKSVEEYMKKE

GGATCCATTGGGTACCCGTTGTAAGTCCGTGGAAGAGTATATGAAGAAGGAAACAC TPHKHLIWLNKCDLVPTWVAV

CACACAAGCATTTAATTTATGTTCTTAATAAGTGTGATTTGGTACCTACCTGGGTTGC C*LFFFFF*FFPTKRIAREIFFFDR

AGTATGTTAAC I I I I I I I I I I I I I I I I I I GATTCTTTCCAACCAAGAGGATAGCACGAG V*FLLPNLFVRVLLYPF^*DKLIYR

AGA I I I I I I I I I I CGATAGAGTATGA I I I I I ATTACCAAACC I I I I I GTCAGGGTGCTT SVCYCFGRVILFAYPYFPVLGKP

CTCTATCCGTTTTAGGATAAACTTATCTACAGAAGTGTCTGTTACTGTTTTGGAAGAG HG^*IRRTTFVLKVYLRFLEQRGS

TAATACTGTTCGCTTATCCATATTTCCCCGTTCTGGGGAAGCCTCATGGGTAAATTAG YELSFCYLRLYLGLHTICS^**RSS

AAGGACAACCTTTGTTTTAAAGGTATACCTTCGCTTTTTAGAACAGCGAGGATCTTAT QFSTFFFFLV^*STLRNNQSRL^*Q*

GAGTTGAGCTTTTGTTATTTGAGACTTTATCTCGGGCTCCATACAATATGTTCGTAGT LLHSLGAKDYLL*NKMYFLLTEG

AAAGATCCTCACAATTTTCTAC I I I I I I I I I I I I I I I AGTTTAATCCACTCTTCGAAACA FFFQF^*LYK*AAWVKHLSKERPT

ATCAATCTCGGCTATAACAATGATTATTACATTCTCTTGGGGCAAAAGATTACCTATT LAFHASITNSFGKGSLIQLLRQF

ATGAAACAAAATGTAI I I I I I ACTAACAGAAGG I I I I I I I I I I CAATTCTAACTTTACAA SQLHTDRKQISVGFIGYPNTGKS

GTAGGCAGCTTGGGTCAAACATTTGTCAAAGGAACGTCCAACTTTGGCGTTTCACGC SIINTLRKKKVCQVAPIPGETKV

ATCTATTACCAACTCTTTCGGTAAAGGTTCGTTAATTCAGTTGCTACGTCAGTTCTCA W

CAGTTGCATACTGATAGAAAGCAAATATCTGTAGGGTTTATCGGCTATCCAAACACT

GGTAAATCGTCCATCATTAACACATTGAGAAAGAAAAAGGTGTGTCAAGTTGCACCA

ATCCCTGGTGAAACTAAGGTCTGGCA

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YDR328C 841 AATTCGATATCTCGCCTTGGTGAGGAAAATTTACCAGGTCTGTCGATTAGTGTAGCC 842 NSISRLGEENLPGLSISVAAEPM

GCTGAACCAATGGACTCTAAAATGATTCAGGATTTGAGCAGGAACACATTGGGAAAG DSKMIQDLSRNTLGKGQNCLDI

GGTCAAAACTGCTTGGATATTGACGGAATAATGGACAACCCAAGGAAACTATCCAAA DGIMDNPRKLSKILRTEYGWDS

ATATTGAGAACCGAGTATGGGTGGGATTCGTTGGCATCAAGAAACGTTTGGTCTΠT LASRNVWSFYNGNVLINDTLPD

TATAATGGCAACGTGTTGATTAATGACACTTTGCCAGATGAAATCAGTCCTGAATTAT EISPELLSKYKEQIIQGFYWAVK

TATCCAAATATAAGGAACAAATAATACAAGGATTTTACTGGGCTGTAAAAGAGGGGC EGPLAEEPIYGVQYKLLSISVPS

CTTTGGCAGAAGAACCAATTTATGGTGTACAGTATAAATTGTTATCGATCTCAGTACC DVNIDVMKSQIIPLMKKACWGL

TTCCGACGTAAACATTGACGTTATGAAAAGTCAAATTATTCCGCTAATGAAAAAAGCC LTAIPILLEPIYEVDITVHAPLLPIV

TGTTACGTTGGCTTACTGACAGCAATCCCAATTTTATTGGAACCCATCTATGAAGTTG EELMKKRRGSRIYKTIKVAGTPL

ACATCACGGTCCACGCCCCCTTGCTGCCAATAGTAGAGGAACTTATGAAGAAGAGA LEVRGQVPVIESAGFETDLRLST

CGTGGAAGCAGGATATACAAAACAATAAAAGTGGCAGGGACACCATTGTTGGAGGT NGLGMCQLYFWHKIWRKVPGD

TCGTGGACAAGTTCCGGTTATTGAATCTGCAGGATTCGAGACAGATTTGAGATTATC VLDKDAFIPKLKPAPINSLSRDFV

TACGAATGGTCTTGGTATGTGTCAGCTGTACTTTTGGCACAAGATATGGAGGAAGGT MKTRRRKGISTGGFMSNDGPTL TCCTGGTGATGTTTTGGATAAAGATGCGTTTATTCCAAAATTGAAACCCGCACCTATC EKYISAELYAQLRENGLVP*KSW AACAGTTTAAGTCGTGATTTCGTGATGAAAACAAGAAGGCGGAAGGGTATTTCTACA RIRIKQWHNIFIQQNLHMTSYLV GGTGGATTTATGTCAAATGATGGTCCTACGCTTGAAAAGTATATAAGCGCTGAATTAT l*LLIGTENFLFLPKYR*DELYRIL ACGCTCAATTAAGAGAAAATGGCTTAGTACCGTGAAAATCTTGGCGCATAAGAATTA RC*KVKR*QSTVS*TTVYREWLK AGCAATATGTCCACAATATTTTTATTCAGCAGAATTTACATATGACTTCGTATTTAGTG VSS*RSCFRIKRKLKKLKSLKMT^* ATATAACTACTCATCGGGACTGAAMTTTTTTATTTCTGCCTAAATACAGATGAGATG RKRNPKN^*RKKNQLSLQLLI^*KSL AGCTTTATAGAATTTTACGCTGTTAAAAAGTTAAACGGTAACAAAGCACAGTTAGTTG RRKKRRPMLRRKLLQILKNIKAK AACAACGGTATATAGAGAATGGCTAAAGGTTTCAAGTTGAAGGAGTTGCTTTCGCAT PFLKKRKES*KKN*KKCKNRTLP CAAAAGGAAATTGAAAAAGCTGAAAAGCTTGAAAATGACCTAAAGAAAAAGAAATCC KLKSICLAMK CAAGAATTGAAGAAAGAAGAACCAACTATCGTTACAGCTTCTAATTTGAAAAAGCTTG AGAAGAAAGAAAAGAAGGCCGATGTTAAGAAGGAAGTTGCTGCAGATACTGAAGAAT ATCAAAGCCAAGCCCTTTCTAAAAAAGAGAAAAGAAAGTTGAAAAAAGAATTGAAAAA AATGCAAGAACAGGACGCTACCGAAGCTCAAAAGCATATGTCTGGCGATGAAG

YDR328C 847 CTTTATCAGGTGGCCGACACTAGGGAATAAGACAGCATGGAGCGGCCTGGCTTGGT 848 LYQVADTRE^*DSMERPGLVLQD

ATTGCAGGACCTTCCACCCGAAATCCTGATAAACATCTTTTCTCACTTAGACGAAAAG LPPEILINIFSHLDEKDLFTLQELS

GACTTGTTCACGTTGCAAGAACTTTCTACGCATTTCCGGAACTTAATTCACGATGAAG THFRNLIHDEELWKNLFKSRVH

AACTTTGGAAGAACCTGTTTAAGTCTAGAGTTCACACGACTCATTTCCCCACATTTTC TTHFPTFSQSSKFSVEYIERTRG

TCAGTCTTCGAAATTTAGTGTAGAGTATATTGAAAGGACACGGGGTCTTCATCATTG LHHWQHNKAIRTKYTIIPTRNWD

GCAGCATAACAAGGCTATAAGAACCAAATACACAATAATTCCGACGAGAAACTGGGA QPSIERIVFDYPRVAAYNDGTITI

TCAGCCCAGTATAGAACGCATAGTGTTTGATTATCCACGAGTTGCAGCTTATAATGAT LQLQNHKRQKKFKKLIYIPCTTP

GGTACCATTACAATTTTACAGTTACAAAATCACAAAAGGCAGAAGAAATTTAAGAAAT QGCSTMDFNINAAVFGRFDGRV

TGATTTATATTCCATGTACAACACCACMGGTTGTTCTACCATGGACTTCAATATTAAT FGKLLSNKSYLTPVMEFTGRHS

GCTGCAGTGTTTGGTAGGTTTGACGGCAGAGTGTTTGGAAAACTGCTAAGTAACAAA AGVTAICNSESWDTSREDWSV

TCGTACTTAACGCCCGTGATGGAATTTACGGGAAGGCATAGTGCAGGCGTTACTGC SGSENGEIIWWCENKLVKMWK

CATTTGTAATTCAGAATCCTGGGACACGTCAAGAGAGGACTGGTCTGTTAGTGGCTC VSNRVIWKLAFFKDWTLIMDDE

TGAAAACGGTGAAATTATATGGTGGTGCGAAAATAAGCTTGTTAAGATGTGGAAGGT KLYIIHQMQELHSIDIPKDLDEQP

TTCTAATAGGGTAATTTGGAAACTGGC I I I I I I I AAAGATTGGACCTTAATTATGGAT MRVRFFKMDFGSMTLVLADLNN

GATGAAAAGCTGTACATTATCCATCAAATGCAGGAATTGCACAGTATTGATATTCCTA VYTISVNPNGNFGNLRKLEMPE

AGGATCTGGACGAACAGCCTATGAGGGTACGGTTTTTCAAGATGGATTTCGGTTCCA QICAVEIDEKTSQREQNWQFAG

TGACTTTGGTACTTGCTGACCTGAATAACGTTTATACAATATCGGTCAATCCCAATGG DDGCYISLLTTQNTLYIINIRDLS

TAACTTCGGCAATTTGAGAAAGTTGGAGATGCCTGAACAAATATGCGCAGTTGAAAT SSGLKVQCKISFDEQVWSQVT

AGATGAAAAAACTTCTCAAAGGGAACAAAACTGGCAATTTGCTGGTGATGACGGCTG NLIVWALPNVLQILNAMTGELIK

CTATATCAGTCTTTTGACGACACAGAACACTTTATACATTATCAATATAAGAGATCTTT TVLKTEKFPEFLKVSQDKIIMGS

CTTCGTCAGGCTTAAAAGTTCAATGTAAGATAAGTTTTGATGAGCAGGTATACGTATC GNVLNYLKFVSSDSKKHHHSTK

GCAGGTGACAAATTTAATAGTTGTGGTAGCATTGCCAAATGTTCTTCAAATATTGAAC GKNTVSNKWNETLNTELQYYDE

GCGATGACAGGCGAGTTAATAAAGACAGTGTTGAAAACCGAGAAGTTTCCCGAATTT D

TTGAAGGTATCGCAGGATAAAATCATTATGGGAAGCGGTAACGTTTTGAATTATTTAA

YDR328C 853 CCAGGAACAGAAGCAGGGATCGTTCCGGTTTCAGCAAACACTCCAAAAAGCTTGAAT 854 PGTEAGIVPVSANTPKSLNSNINI

AGCAATATTAATATCAACGTAAATAATAACAATATTGGCCAACAGCAAGTTAAGAAGC NVNNNNIGQQQVKKPRKQRVK

CAAGAAAGCAAAGAGTGAAAAAAAAGACCAAAAAGGAATTGGAACTAGAACGTAAAG KKTKKELELERKEREDFQKRQQ

AMGGGAGGATTTTCAGAAACGACAACAAAAACTTTTAGAGGATCAACAAAGGCAAC KLLEDQQRQQKLLLETKLRQQY

AGAAATTGCTATTAGAGACAAAATTACGTCAACAATATGAAATCGAACTAAAAAAATT EIELKKLPKVYKRSIVRNYKPLIN

GCCTAAAGTCTACAAGAGATCAATTGTTAGGAACTACAAACCCCTAATCAACCGCCT RLKHYNGYDINYISKIGEKIDSNK

CAAGCATTACAATGGTTACGATATCAATTACATCTCTAAAATAGGAGAGAAAATAGAT PIFLFAPELGAINLHALSMSLQSK

TCCAACAAGCCAA I I I I I CTCTTCGCGCCAGAGTTAGGTGCAATTAATTTACATGCTT NLGEINTALNTLLVTSADSNLKIS

TATCAATGTCCCTCCAATCGAAGAATCTTGGAGAAATAAACACCGCCTTGAACACCTT LVKYPELLDSLAILGMNLLSNLS

GTTGGTCACAAGCGCTGACTCGAACTTAAAAATATCTCTGGTCAAATACCCTGAATTA QNWPYHRNTSDYYYEDAGSN

TTAGACTCCTTGGCAATACTCGGCATGAATTTACTGTCAAATTTGTCACAAAATGTTG QYYVTQHDKMVDKIFEKVNNNA

TTCCATACCATCGAAACACTTCTGACTATTATTATGAGGATGCTGGATCAAATCAATA TLTPNDSNDEKVTILVDSLTGNQ

CTATGTTACCCAACACGATAAAATGGTTGATAAAATTTTTGAAAAGGTAAACAACAAC LPTPTPTEMEPDLDTECFISMQS

GCTACACTTACACCGAATGATTCTAACGATGAAAAAGTCACTATCCTGGTAGATTCTT TSPAVKQWDLLPEPIRFLPNQF

TAACAGGTAATCAATTGCCCACCCCTACTCCTACTGAAATGGAGCCTGATCTCGACA PLKIHRTPYLTSLKKIKDEIDDPF

CTGAATGTTTTATAAGTATGCAGTCGACATCTCCCGCAGTTAAACAGTGGGACTTATT TKINTRGAEDPKVLINDQLSTIS

GCCTGAACCAATAAGATTCCTCCCTAACCAATTTCCTCTGAAAATTCACAGAACTCCT MILRNISFSDNNSRIMSRNFYLK

TATTTGACTTCTTTGAAAAAAATCAAGGATGAAATTGATGATCCATTTACAAAAATAAA RFISDLLWLVLIHPENFTCNRKIL

TACCAGAGGGGCAGAGGATCCCAAAGTTCTGATTAACGATCAACTGTCTACCATCTC NFKKDLVIVLSNISHLLEIASSIDC

GATGATTTTGAGGAATATTTCATTCTCCGATAACAACTCCAGAATCATGTCGAGAAAT LLILILVISFGQPKLNPMASSSSF

TTTTACCTAAAGAGATTTATATCTGATCTACTTTGGTTAGTCTTAATCCATCCAGAAAA GSESLTFNEFQLQWGKYQTFG

CTTTACATGCAATAGGAAAATACTA TTTCAAGAAGGATTTGGTTATTGTTTTATCAA VDILAKLFSLEKPNLNYFKSILLN

ATATTTCTCATTTATTAGAGATCGCTTCGTCCATTGATTGCTTGTTAATTCTTATTCTA KNTGNNLYDRNSNNNHKDKKLL

GTCATAAGTTTTGGTCAACCAAAACTCAATCCAATGGCGTCTTCGTCATCATTTGGCT R

Claims

What is claimed is:

1. A complex between two polypeptides of Saccharomyces cerevisiae as set forth in columns 1 and 2 of Table I, respectively.

2. A complex between two polynucleotides of Saccharomyces cerevisiae , said polynucleotides encoding two polypeptides as set forth in columns 1 and 2 of Table I, respectively.

3. A recombinant host cell expressing a polynucleotide encoding a Saccharomyces cerevisiae polypeptide as set forth in columns 1 and 2 of Table I.

4. A method for selecting a modulating compound that inhibits or activates the interaction between two polypeptides of Saccharomyces cerevisiae comprising:

(a) cultivating a recombinant host cell comprising a reporter gene on a selective medium containing a modulating compound the expression of which is toxic for said recombinant host cell wherein said recombinant host cell is transformed with two vectors:

(i) wherein said first vector comprises a polynucleotide encoding a first hybrid polypeptide and a DNA bonding domain; (ii) wherein said second vector comprises a polynucleotide encoding a second hybrid polypeptide and an activating domain that activates said toxic reporter gene when the first and second hybrid polypeptides interact;

(b) selecting said modulating compound which inhibits the growth of said recombinant host cell.

5. A modulating compound obtained from the method of Claim 4.

6. A SID® polypeptide comprising the even SEQ ID Nos. from 2 to 864 (column 3 of Table II).

7. A SID® polynucleotide comprising the uneven SEQ ID Nos. from 1 to 863 (column 2 of Table II).

8. A vector comprising a SID® polynucleotide comprising the uneven SEQ ID Nos. from

1 to 863 (column 2 of Table II).

9. A fragment of said SID® polypeptide according to Claim 6.

10. A variant of said SID® polypeptide according to Claim 6.

11. A fragment of said SID® polynucleotide according to Claim 7.

12. A variant of said SID® polynucleotide according to Claim 7.

13. vector comprising the SID® polynucleotide according to Claim 1 1 or 12.

14. A recombinant host cell containing the vectors according to Claim 8 or 13.

15. A pharmaceutical composition comprising the modulating compound of Claim 5 and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising the SID® polypeptide according to Claim 6 and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising the recombinant host cells of Claim 14 and a pharmaceutically acceptable carrier.

18. A protein chip comprising the polypeptides according to Claim 6, 9 or 10.