Furthermore, the present invention generally relates to the identification of genes associated with infection by identifying genes that are regulated in terms of expression in the course of an infection.

Several documents are cited throughout the text of this specification. Each of the documents cited herein (including any manufacturer's specifications, instructions, etc) are hereby incorporated by reference ; however, there is no admission that any document, cited is indeed prior art of the present invention.

The Epstein-Barr virus is a ubiquitous human herpesvirus that is associated with an increasing number of human malignancies. Epstein-Barr virus (EBV) infects almost the entire adult population of the world (Faulkner, Trends Microbiol. 8 (2000), 185-189). The success of this virus appears to be based on its ability to infect the B cell, rather than any other cell type. Among human malignancies are Epstein-Barr virus-associated lymphoproliferative diseases in immuno- compromised patients, a spectrum of mainly B-cel ! diseases that range from polyclonal lymphoproliferative diseases, which resolve when immunosuppression is halted, to highly malignant lymphomas (Mosier, Curr. Opin. Hemato. 6 (1999), 25-29). Thus, it has been known for 30 years that EBV is the etiologic agent of acute infectious mononucleosis and is closely associated with the genesis of Burkitt's lymphom and undifferentiated nasopharyngeal carcinoma. Recent studies have demonstrated that EBV is also implicated in a variety of other diseases, such as EBV-associated hemophagocytic syndrome, chronic active EBV infection, T-cell lymphoma, natural killer cell leukemia/lymphoma, lymphoproliferative diseases in immunocompromised hosts, Hodgkin's disease, pyothorax-associated B-cell lymphom, smooth-muscle tumors, and gastric carcinoma (for review see Kawa, Int. J. Hematol. 71 (2000), 108-117). Until recently, there has been no definite treatment for severe EBV infection but many therapeutic approaches have been attempted, and some of them seem to be beneficial for a certain EBV-associated disease (Okano, Pediatr. Hemato. Oncol.

14 (1997), 109-119). However, there remains a constant need for targets and strategies for the treatment of EBV infection and diseases related to EBV infection.

Thus, the technical problem underlying the present invention is to provide methods and means for the development of additional effective therapies for EBV infected humans and animals. The solution to this technical problem is achieved by providing the embodiments characterized in the claims.

Accordingly, the present invention relates to a method for identifying an antagonist or inhibitor of EBV infection, said method comprising the steps of (a) testing a candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors for the inhibition or reduction of transcription of a gene encoding a polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof; or (b) testing a candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors for the inhibition or reduction of translation of mRNA transcribed from said gene or a fragment or derivative or ortholog thereof; and (c) determining whether said candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors tests positive in step (a) and/or (b).

The present invention arose from experiments based on a conditional cellular system for Epstein-Barr virus nuclear antigen 2 (EBNA2). Epstein-Barr virus (EBV) infects and immortalizes primary human B-cells very efficiently. Epstein- Barr virus nuclear antigen 2 (EBNA2) is one of the first viral genes expressed post-infection. EBNA2 is a transactivator of cellular and viral target genes, which carries an intrinsic transactivation domain but cannot bind to DNA directly. It gains access to the promoter of target genes via DNA binding anchor proteins. A conditional cellular system for EBNA2 (the ER/EB cell line) has been generated and used for functional studies in EBV immortalized B-cells. It could be shown that EBNA2 induces and maintains the proliferation of EBV'immortalized B-cells.

EBNA2 directly activates the transcription of master genes, like the viral LMP1 and the cellular c-myc gene. Activation of these master genes initiates a cascade of events, which indirectly induces the expression of genes relevant for cell cycle progression like cdk4.

In accordance with the present invention expression profiling for a systematic approach towards the identification of novel EBNA2 target genes has been employed for novel EBNA2 target genes. To this end time course experiments were performed in order to follow the induction of genes post-EBNA2 activation in ER/EB cells. ExpressCode (TM) technology has been applied : 9,000 cDNA clones were arrayed at high densities on the 8 x 12 cm nylon membrane and hybridized with 33P-labelled cDNA transcribed from RNA populations of different time points post-stimulation. Statistical confidence of the data was derived by 4 replicates of each experiment. By hierarchical clustering of the results, group specific expression profiles for a subset of genes could be identified. Thus, four genes could be identified for the first time to be associated with the expression of an EBV encoded gene product and thus are expected to be relevant for EBV infection. These identified genes encode TRRAP, ATF5, elF1 and SUPT6H, respectively, which are further discussed below.

TRRAP: One of the identified genes upregulated upon EBNA2 is the ATM-related protein TRRAP (GenBank accession number NM003496) which is an essential cofactor for the c-Myc and E2F oncoproteins (McMahon, Cell 94 (1998), 363-374; Vassilev, Mol. Cell. 2 (1998), 869-875). The c-Myc and E2F transcription factors are among the most potent regulators of cell cycle progression in higher eukaryotes. The highly conserved 434 kDa protein, designated TRRAP, interacts specifically with the c-Myc N terminus and has homology to the ATM/P13-kinase family. TRRAP also interacts specifically with the E2F-1 transactivation domain.

Expression of transdominant mutants of the TRRAP protein or antisense RNA blocks c-Myc-and E1A-mediated oncogenic transformation. Such mutants and antisense RNA can of course be used in accordance with the present invention to prevent, delay and/or treat EBV infection or provide a lead for improved antagonists and inhibitors. Since TRRAP does not seem to be essential for cellular growth TRRAP is particularly preferred as a target. Furthermore, c-myc antagonists and inhibitors may be used in combination with those of TRRAP.

ATF5: The human activating transcription factor 5 (ATF5) encoding nucleotide sequence can be retrieved from Genbank accession number NM012068 ; see also Pati, Mol. Cell. Biol. 19 (1999), 5001-5013. ATF5 has been discovered with a yeast-based genetic assay designed to identify proteins that interact with human Cdc34. Such binding assays can of course also be used to identify antagonists and inhibitors of ATF5. Approaches to identify antagonists and inhibitors of ATF5 can be adapted from the prior art. For example, the isolation of a dominant transcription inhibitor in ATF-a heterodimers has been described (Pescini, J. Biol.

Chem. 269 (1994), 1159-1165). Furthermore, the use of an anti-ATF-1 single chain Fv fragment as inhibitor has been described (Jean, Oncogene 19 (2000), 2721-2730). elF1 : The cloning and characterization of the human translation initiation factor 1 (elF1 (A121/SU11)) encoding gene (GenBank accession number NM005801) is described in Sheikh, J. Biol. Chem. 274 (1999), 16487-16493; Lian, Oncogene 18 (1999), 1677-1687; Fields and Adams, Biochem. Biophys. Res. Commun. 198 (1994), 288-291. The full-length human cDNA contains an open reading frame of 113 amino acids, corresponding to a protein of 12.7 kDa. The deduced amino acid sequence of shows high homology to the yeast translation initiation factor (elF) sui1. Expression of human A121 corrected the mutant sui1 phenotype in yeast, demonstrating that human A121 encodes a bona fide translation initiation factor that is equivalent to yeast suilp. Accordingly, this assay, e. g. the mutant sui1 yeast complemented with human elF1, can conveniently be used to screen for antagonists and inhibitors of elF1 in accordance with the methods of the present invention described herein. Furthermore, the structure of this factor can be determined, for example as described for the elF1A solution structure that revealed a large RNA-binding surface important for scanning function (Battiste, Mol. Cell. 5 (2000), 109-119 and Fletcher, EMBO J. 18 (1999), 2631-2637). Such structures can then be used for computer assisted drug design, for example according to the methods described below.

SUPT6H: The identification and analysis of the human and murine putative chromatin structure regulator SUPT6H (GenBank accession number U46691) and Supt6h is described in Chiang, Genomics 34 (1996), 328-333 and Genomics 38 (1996), 421-424. SUPT6H and Supt6h are the human and murine homologues of the Saccharomyces cerevisiae and Caenorhabditis elegans genes SPT6 (P using 1603 aa = 6.7 e-95) and emb-5 (P using 1603 aa = 7.0 e-288), respectively. The human and murine SPT6 homologues are virtually identical, as they share >98% identity and >99% similarity at the protein level. The derived amino acid sequences of these two genes predict a 1603-aa protein (human) and a 1726-bp protein (mouse), respectively. There were several known features, including a highly acidic 5'-region, a degenerate SH2 domain, and a leucine zipper. These features are consistent with a nuclear protein that regulates transcription, whose extreme conservation underscores the likely importance of this gene in mammalian development. Since SUPT6H finds its homologues in Saccharomyces cerevisiae and Caenorhabditis elegans corresponding model systems can be used for screening antagonists and inhibitors in accordance with the present invention. The set up of such screening systems is well within the skill of the person skilled in the art; for example see, e. g., Parissi, Gene 247 (2000), 129-136.

DNA sequences encoding the above mentioned proteins and corresponding amino acid sequences are also provided in Table I.

Since the described genes have been shown to be specifically associated with EBNA2 expression it is expected that antagonizing and/or inhibiting these genes or the activity of their encoded products, at least to a level which substantially corresponds to the expression of the gene or protein function in a non-infected cell, will give raise to a reduced EBV infection, for example in terms of viral replication or spreading, or with respect to severity of disease symptoms, for example of diseases and disorders related to EBV infection.

Thus, an antagonist or inhibitor of the expression of any one of the above identified genes or gene products either alone or in combination provides a further means for a therapy of EBV infection. Thus, the method of the present invention provides the option of development of new anti EBV agents. The findings of the present invention are particularly important in view of the drawbacks of the present forms of treatment of EBV infections, and diseases and disorders related to EBV infections.

The term"antagonist"or"inhibitor"as used herein means naturally occurring and synthetic compounds capable of counteracting or inhibiting an activity of a gene or gene product or interactions of the gene or gene product with other genes or gene products. Determining whether a compound is capable of inhibiting or counteracting specific gene expression can be done, for example, by Northern blot analysis, Western blot analysis or proteome analysis. It can further be done by monitoring the phenotypic characteristics of a bacterial cell contacted with the compounds and compare it to that of a wild-type cell. In an additional embodiment, said characteristics may be compared to that of a cell contacted with a compound which is either known to be capable or incapable of suppressing or activating the protein or gene, respectively, according to the invention. For example, a transgenic cell can be used and the phenotypic characteristics comprises a readout system. Further examples of determining whether a compound is capable of inhibiting or counteracting specific gene expression are described below.

The term"expression"means the production of a protein or nucleotide sequence in a cell. However, said term also includes expression of the protein in a cell-free system. It includes transcription into an RNA product, and/or translation into a polypeptide from a DNA encoding that product.

The term"transcription"as used herein means a DNA template dependent synthesis of a ribonucleic acid polymer encoding a polypeptide or a regulatory sequence. The term"translation"as used herein means the polymerization of a polypeptide that is encoded by an RNA molecule by a protein complex.

As used in accordance with the present invention, the term"fragment or derivative"denotes any variant the amino acid or nucleotide sequence of which deviates in its primary structure, e. g., in sequence composition or in length as well as to analogue components. For example, one or more amino acids of a polypeptide may be replaced in said fragment or derivative as long as the modified polypeptides remain functionally equivalent to their described counterparts. The term"fragment or derivative"further denotes compounds analog to an antagonist or inhibitor that should have a stabilized electronic configuration and molecular conformation that allows key functional groups to be presented to the mentioned polypeptide in substantially the same way as the antagonist and inhibitor. The variant of the polypeptide may be a naturally occurring allelic variant of the polypeptide or non-naturally occurring variants of those polynucleotides.

The term"orthologs"as used herein means homologous sequences in different species that evolved from a common ancestral gene by speciation. Normally, orthologs retain the same function in the course of evolution. However, orthologous genes may or may not be responsible for a similar function (see, e. g., the glossary of the"Trends Guide to Bioinformatics", Trends Supplement 1998, Elsevier Science). Orthologous genes, nucleic acids or proteins comprise genes, nucleic acids or proteins which have one or more sequences or structural motifs in common. For example, the sequence motifs of proteins can comprise short, i. e. repetitive sequences or amino acid positions conserved in the primary structure and/or conserved in higher protein structures, e. g. secondary or tertiary structure.

Orthologous nucleic acids or genes can comprise molecules having short stretches of one or more homologous (same or similar) sequences, for example protein binding boxes or structure forming boxes. Methods for the identification of a candidate ortholog of a gene or polypeptide described herein are known to those skilled in the art and are described for example in Sambrook et al. (1989), Molecular Cloning : A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York, or Ausubel (1994), Current Protocols in Mol. Biol.. The person skilled in the art knows how to identify orthologous genes, nucleic acids or polypeptides by computer supported analysis (e. g. BLAST) of known sequences and its interpretation.

The terms"gene","polynucleotide","nucleic acid sequence","nucleotide sequence","DNA sequence"or"riucleic acid molecule"as used herein refer to polymeric forms of nucleotides of any length, either ribonucleotides or deoxyribonucleotides and only to the primary structure of the molecule. Thus, these terms include double-and single-stranded DNA, and RNA. They also include known types of modifications, for example, methylation,"caps" substitution of one or more of the naturally occurring nucleotides with an analog.

Preferably, the DNA sequence of the invention comprises a coding sequence encoding at least the mature form of the above defined protein, i. e. the protein which is posttranslationally processed in its biologically active form, for example due to cleavage of leader or secretory sequences or a proprotein sequence or other natural proteolytic cleavage points.

The term"plurality of candidate antagonists or inhibitors"is to be understood as a plurality of substances which may or may not be identical.

Said antagonists or inhibitors or plurality of candidate antagonists or inhibitors may be chemically synthesized or microbiologically produced and/or comprised in, for example, samples, e. g., cell extracts from, e. g., plants, animals or microorganisms. Furthermore, said compound (s) may be known in the art but hitherto not known to be capable of suppressing or inhibiting said polypeptide.

The reaction mixture may be a cell free extract or may comprise a cell or tissue culture. Suitable set ups for the method of the invention are known to the person skilled in the art and are, for example, generally described in Alberts et al., Molecular Biology of the Cell, third edition (1994), in particular Chapter 17. The plurality of compounds may be, e. g., added to the reaction mixture, culture medium, injected into the cell or sprayed onto the plant.

In line with the above, the present invention also relates to a method for testing a candidate EBV antiviral agent comprising the steps of (a) contacting a polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof with a candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors; and (b) determining whether said antagonist or inhibitor binds to said polypeptide.

For example, a candidate antagonist or inhibitor not known to be capable of binding to an polypeptide encoded by a essential gene as described above can be tested to bind thereto comprising contacting a said polypeptide with a candidate antagonist or inhibitor under conditions permitting binding of ligands known to bind thereto, detecting the presence of any bound ligand, and thereby determining whether such candidate antagonist or inhibitor inhibits the binding of a ligand to a polypeptide as described above.

Proteins that bind to a polypeptide as described above and might inhibit or counteract to said polypeptide can be"captured"using the yeast two-hybrid system (Fields, Nature 340 (1989), 245-246). A modified version of the yeast two- hybrid system has been described by Roger Brent and his colleagues (Gyuris, Cell 75 (1993), 791-803; Zervos, Cell 72 (1993), 223-232). Briefly, a domain of the polypeptide is used as bait for binding compounds. Positives are then selected by their ability to grow on plates lacking leucine, and then further tested for their ability to turn blue on plates with X-gal, as previously described in great detail (Gyuris, supra; WO 95/31544). Once amino acid sequences are identified which bind to a polypeptide encoded by a gene selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof, these sequences can be screened for antagonist activity using, for example, an proliferation assay or used for screening for antagonists of said binding.

Another assay which can be performed to identify inhibitors and antagonists involves the use of combinatorial chemistry to produce random peptides which then can be screened for both binding affinity and antagonist effects. One such assay has recently been performed using random peptides expressed on the surface of a bacteriophage (Wu (1996), Nature Biotechnology 14,429-431).

In a preferred embodiment of the method of the present invention said method further comprises identifying an antagonist or inhibitor optionally from said sample of candidate antagonists or inhibitors.

If a sample contains a candidate antagonist or inhibitor, or a plurality of candidate antagonists or inhibitors, as identified in the method of the invention, then it is either possible to isolate the candidate antagonists or inhibitors from the original sample identified as containing the compound capable of binding to anyone of the above described polypeptides or antagonizing their activity in a cellular assay, or one can further subdivide the original sample, for example, if it consists of a plurality of different candidate antagonists or inhibitors, so as to reduce the number of different substances per sample and repeat the method with the subdivisions of the original sample. Depending on the complexity of the samples, the steps described above can be performed several times, preferably until the sample identified according to the method of the invention only comprises a limited number of or only one substance (s). Preferably said sample comprises substances of similar chemical and/or physical properties, and most preferably said substances are identical. As regards the identification of candidate antagonists or inhibitors by any of the above-referenced embodiments of the invention, a variety of formats or tools is available to the person skilled in the art.

Thus, several methods are known to the person skilled in the art for producing and screening large libraries to identify compounds having specific affinity for a target. These methods include the phage-display method in which randomized peptides are displayed from phage and screened by affinity chromatography to an immobilized receptor; see, e. g., WO 91/17271, WO 92/01047, US-A-5,223,409. In another approach, combinatorial libraries of polymers immobilized on a chip are synthesized using photolithography ; see, e. g., US-A-5,143,854, WO 90/15070 and WO 92/10092. The immobilized polymers are contacted with a labeled polypeptide and scanned for label to identify polymers binding to the polypeptide.

The synthesis and screening of peptide libraries on continuous cellulose membrane supports that can be used for identifying binding ligands of the polypeptide to be used in accordance with the invention and thus possible inhibitors and antagonists is described, for example, in Kramer, Methods Mol.

Biol. 87 (1998), 25-39. This method can also be used, for example, for determining the binding sites and the recognition motifs in the polypeptide as described above. In like manner, the substrate specificity of the DnaK chaperon was determined and the contact sites between human interleukin-6 and its receptor ; see Rudiger, EMBO J. 16 (1997), 1501-1507 and Weiergraber, FEBS Lett. 379 (1996), 122-126, respectively. Furthermore, the above-mentioned methods can be used for the construction of binding supertopes derived from the described polypeptides. A similar approach was successfully described for peptide antigens of the anti-p24 (HIV-1) monoclonal antibody; see Kramer, Cell 91 (1-997), 799-809. A general route to fingerprint analyses of peptide-antibody interactions using the clustered amino acid peptide library was described in Kramer, Mol. Immunol. 32 (1995), 459-465. In addition, antagonists or inhibitors of a polypeptide described above can be derived and identified from monoclonal antibodies that specifically react with any one of said polypeptides in accordance with the methods as described in Doring, Mol. Immunol. 31 (1994), 1059-1067.

More recently, WO 98/25146 described further methods for screening libraries of complexes for compounds having a desired property, especially, the capacity to agonize, bind to, or antagonize a polypeptide or its cellular receptor. The complexes in such libraries comprise a compound under test, a tag recording at least one step in synthesis of the compound, and a tether susceptible to modification by a reporter molecule. Modification of the tether is used to signify that a complex contains a compound having a desired property. The tag can be decoded to reveal at least one step in the synthesis of such a compound. Other methods for identifying compounds which interact with the proteins according to the invention or nucleic acid molecules encoding such molecules are, for example, the in vitro screening with the phage display system as well as filter binding assays or "real time"measuring of interaction using, for example, the BlAcore apparatus (Pharmacia).

All these methods can be used in accordance with the present invention to identify antagonists and inhibitors of a polypeptide encoded by a gene selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof.

Additionally, the present invention relates in a preferred embodiment to a method comprising improving inhibitors or antagonists identified by peptidomimetics or by applying phage display or combinatorial library technique step (s).

Peptidomimetics, phage display and combinatorial library techniques are well- known in the art and can be applied by the person skilled in the art without further ado to the improvement of the antagonist or inhibitor that is identified by the basic method referred to herein above.

Methods for the generation and use of peptidomimetic combinatorial libraries are described in the prior art, for example in Ostresh, Methods In Enzymology 267 (1996), 220-236; Dosner, Bioorg. Med. Chem. 4 (1996), 709-715; Beeley, Trends Biotechn. 12 (1994), 213-216; al-Obeidi, Mol. Biotechn. 9 (1998), 205-223 ; Wiley, Med. Res. Rev. 13 (1993), 327-384; Bohm, J. Comput. Aided Mol. Des. 10 (1996), 265-272 ; and Hruby, Biopolymers 43 (1997), 219-266.

Various sources for the basic structure of such an antagonist or inhibitor can be employed and comprise, for example, mimetic analogs of the above described polypeptides or analogs of antagonists or inhibitors already known but hitherto used for a different purpose. Mimetic analogs of the polypeptide of the invention or biologically active fragments thereof can be generated by, for example, substituting the amino acids that are expected to be essential for the biological activity with, e. g., stereoisomers, i. e. D-amino acids; see e. g., Tsukida, J. Med.

Chem. 40 (1997), 3534-3541. Furthermore, in case fragments are used for the design of biologically active analogs pro-mimetic components can be incorporated into a peptide to reestablish at least some of the conformational properties that may have been lost upon removal of part of the original polypeptide ; see, e. g., Nachman, Regul. Pept. 57 (1995), 359-370. Furthermore, the polypeptide can be used to identify synthetic chemical peptide mimetics that bind to or can function as a ligand, substrate, binding partner or the receptor of the polypeptide as effectively as does the natural polypeptide ; see, e. g., Engleman, J. Clin. Invest. 99 (1997), 2284-2292.

The structure-based design and synthesis of low-molecular-weight synthetic molecules that mimic the activity of the native biological polypeptide is further described in, e. g., Dowd, Nature Biotechnol. 16 (1998), 190-195; Kieber-Emmons, Current Opinion Biotechnol. 8 (1997), 435-441; Moore, Proc. West Pharmacol.

Soc. 40 (1997), 115-119; Mathews, Proc. West Pharmacol. Soc. 40 (1997), 121- 125; Mukhija, European J. Biochem. 254 (1998), 433-438.

It is also well known to the person skilled in the art, that it is possible to design, synthesize and evaluate mimetics of small organic compounds that, for example, can act as a substrate or ligand to a polypeptide as identified above. For example, it has been described that D-glucose mimetics of hapalosin exhibited similar efficiency as hapalosin in antagonizing multidrug resistance assistance- associated protein in cytotoxicity; see Dinh, J. Med. Chem. 41 (1998), 981-987.

The gene encoding the polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof or the corresponding RNA can also serve as a target for antagonists or inhibitors.

Antagonists may comprise, for example, proteins that bind to the mRNA of said gene, thereby destabilizing the native conformation of the mRNA and disturbing transcription and/or translation. Furthermore, methods are described in the literature for identifying nucleic acid molecules such as an RNA fragment that mimics the structure of a defined or undefined target RNA molecule to which a compound binds inside of a cell resulting in retardation of cell growth or cell death; see, e. g., WO 98/18947 and references cited therein. These nucleic acid molecules can be used for identifying unknown compounds of pharmaceutical and/or agricultural interest, and for identifying unknown RNA targets for use in treating a disease. These methods and compositions can be used in screening for novel antiviral agents or modifications thereof or for identifying compounds useful to alter expression levels of proteins encoded by a nucleic acid molecule.

Alternatively, for example, the conformational structure of the RNA fragment which mimics the binding site can be employed in rational drug design to modify known antibiotics to make them bind more avidly to the target. One such methodology is nuclear magnetic resonance (NMR), which is useful to identify drug and RNA conformational structures. Still other methods are, for example, the drug design methods as described in WO 95/35367, US-A-5,322,933, where the crystal structure of the RNA fragment can be deduced and computer programs are utilized to design novel binding compounds which can act as inhibitors.

The candidate antagonists and inhibitors which can be tested and identified according to a method of the invention may be taken from expression libraries, e. g., cDNA expression libraries, peptides, proteins, nucleic acids, antibodies, small organic compounds, hormones, peptidomimetics, PNAs or the like (Milner, Nature Medicine 1 (1995), 879-880; Hupp, Cell 83 (1995), 237-245; Gibbs, Cell 79 (1994), 193-198 and references cited supra). Furthermore, genes encoding a putative regulator of a gene encoding the polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof and/or which exert their effects up-or downstream said protein may be identified using, for example, insertion mutagenesis using, for example, gene targeting vectors known in the art (see, e. g., for review West, Med. Res.

Rev. 20 (2000), 216-230 and as an example for conditional gene targeting Takeda, J. Dermatol. Sci. 23 (2000), 147-154). Said compounds can also be functional derivatives or analogues of known inhibitors or antagonists. Such useful compounds can be for example transacting factors which bind an above- described polypeptide. Identification of transacting factors can be carried out using standard methods in the art (see, e. g., Sambrook, supra, and Ausubel, supra). To determine whether a protein binds to the protein or regulatory sequence of the invention, standard native gel-shift analyses can be carried out.

In order to identify a transacting factor which binds to the protein or regulatory sequence of the invention, the protein or regulatory sequence of the invention can be used as an affinity reagent in standard protein purification methods, or as a probe for screening an expression library. The identification of nucleic acid molecules which encode proteins which interact with the polypeptide described above can also be achieved, for example, as described in Scofield (Science 274 (1996), 2063-2065) by use of the so-called yeast"two-hybrid system" ; see also the appended example. In this system, e. g., the protein encoded by the nucleic acid molecules identified in this invention or a smaller part thereof is linked to the DNA- binding domain of the GAL4 transcription factor. A yeast strain expressing this fusion gene and comprising a lacZ reporter gene driven by an appropriate promoter, which is recognized by the GAL4 or LexA transcription factor, is transformed with a library of cDNAs which will express plant genes or fragments thereof fused to an activation domain. Thus, if a peptide encoded by one of the cDNAs is able to interact with the fusion peptide comprising a peptide of a protein of the invention, the complex is able to direct expression of the reporter gene. In this way the nucleic acid molecules and the encoded peptide can be used to identify peptides and proteins interacting with the polypeptide described above. It is apparent to the person skilled in the art that this and similar systems may then further be exploited for the identification of inhibitors or antagonists of the polypeptide.

Once the transacting factor is identified, modulation of its binding to or regulation of expression of the polypeptide described above can be pursued, beginning with, for example, screening for inhibitors against the binding of the transacting factor to the protein specified in accordance with the present invention. Inhibition of EBV infection could then be achieved by applying the transacting factor (or its inhibitor). In addition, if the active form of the transacting factor is a dimer, dominant-negative mutants of the transacting factor could be made in order to inhibit its activity.

Thus, the present invention also relates to the use of a polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof or nucleic acid molecule encoding said polypeptide for the identification of an antagonist or inhibitor of EBV infection.

In another embodiment, the present invention relates to a method for designing an improved antagonist or inhibitor for the treatment of EBV infection or a disorder or disease related to EBV infection comprising the steps of (a) identification of the binding site of an antagonist or inhibitor to the polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof, or identified according to the method of the present invention, by site-directed mutagenesis and chimeric polypeptide studies; (b) molecular modeling of both the binding site of said antagonist or inhibitor and the structure of said polypeptide, and (c) modification of said antagonist or inhibitor to improve its binding specificity or affinity for the polypeptide.

Biological assays as described above or other assays such as assays based on crystallography or NMR may be employed to assess the specificity or potency of the antagonist or inhibitor wherein the decrease of one or more activities of the polypeptide may be used to monitor said specificity or potency. All techniques employed in the various steps of the method of the invention are conventional or can be derived by the person skilled in the art from conventional techniques without further ado.

For example, identification of the binding site of said antagonist or inhibitor by site- directed mutagenesis and chimerical protein studies can be achieved by modifications in the (poly) peptide primary sequence that affect the antagonist's or inhibitor's affinity; this usually allows to precisely map the binding pocket for the drug. Identification of binding sites may be assisted by computer programs. Thus, appropriate computer programs can be used for the identification of interactive sites of a putative antagonist or inhibitor and the polypeptide of the invention by computer assisted searches for complementary structural motifs (Fassina, Immunomethods 5 (1994), 114-120).

As regards step (b), the following protocols may be envisaged: Once the effector site for antagonists or inhibitors has been mapped, the precise residues interacting with different parts of the antagonists or inhibitors can be identified by combination of the information obtained from mutagenesis studies (step (a)) and computer simulations of the structure of the binding site provided that the precise three-dimensional structure of the antagonists or inhibitors is known (if not, it can be predicted by computational simulation). If said antagonist or inhibitor is itself a peptide, it can be also mutated to determine which residues interact with others in the above-mentioned polypeptide essential for EBV replication or spreading.

Finally, in step (c) the antagonist or inhibitor can be modified to improve its binding affinity or its potency and specificity. If, for instance, there are electrostatic interactions between a particular residue of an polypeptide as defined above and some region of an antagonist or inhibitor molecule, the overall charge in that region can be modified to increase that particular interaction. Furthermore, the three-dimensional and/or crystallographic structure of inhibitors or antagonists of the polypeptide of the invention can be used for the design of peptidomimetic inhibitors or antagonists, e. g. in combination with said polypeptide (Rose, Biochemistry 35 (1996), 12933-12944; Rutenber, Bioorg. Med. Chem. 4 (1996), 1545-1558).

Potential antagonists/inhibitors include antisense molecules. Antisense technology can be used to control gene expression through antisense DNA or through triple-helix formation. Antisense techniques are discussed, for example, in Okano, J. Neurochem. 56 (1991), 560; Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, FL (1988). Triple helix formation is discussed in, for instance, Lee, Nucl. Acids Res. 6 (1979), 3073; Cooney, Science 241 (1988), 456; and Dervan, Science 251 (1991), 1360. The methods are based on binding of a polynucleotide to a complementary DNA or RNA.

For example, the 5'coding portion of a polynucleotide that encodes the mature polypeptide as described above may be used to design an antisense RNA oligonucleotide of from about 10 to 40 base pairs in length. A DNA oligonucleotide is designed to be complementary to a region of the gene involved in transcription thereby preventing transcription and the production of the protein. The antisense RNA oligonucleotide hybridizes to the mRNA and blocks translation of the mRNA molecule into receptor polypeptide. As indicated, antagonist or inhibitor e. g. polyclonal and monoclonal antibody according to the teachings of the present invention can be raised according to the methods disclosed in Tartaglia, J. Biol.

Chem. 267 (1992), 4304-4307; Tartaglia, Cell 73 (1993), 213-216, and PCT Application WO 94/09137.

Antibodies may be prepared by any of a variety of methods using immunogens of the polypeptide described above. As indicated, such immunogens include the full length polypeptide (which may or may not include the leader sequence) and fragments such as the ligand binding domain, the extracellular domain and the intracellular domain. These antibodies can be monoclonal antibodies, polyclonal antibodies or synthetic antibodies as well as fragments of antibodies, such as Fab+, Fv, F (ab') 2, disulphide-bridged Fv or scFv fragments, etc. Monoclonal antibodies can be prepared, for example, by the techniques as originally described in Köhler and Milstein, Nature 256 (1975), 495, and Gaufre, Meth.

Enzymol. 73 (1981), 3, which comprise the fusion of mouse myeloma cells to spleen cells derived from immunized mammals. Furthermore, antibodies or fragments thereof to the aforementioned peptides can be obtained by using methods which are described, e. g., in Harlow and Lane"Antibodies, A Laboratory Manual", CSH Press, Cold Spring Harbor, 1988.

The antagonists or inhibitors isolated by the above methods also serve as lead compounds for the development of analog compounds. The analogs should have a stabilized electronic configuration and molecular conformation that allows key functional groups to be presented to the receptor in substantially the same way as the lead compound. In particular, the analog compounds have spatial electronic properties which are comparable to the binding region, but can be smaller molecules than the lead compound, frequently having a molecular weight below about 2 kD and preferably below about 1 kD. Identification of analog compounds can be performed through use of techniques such as self-consistent field (SCF) analysis, configuration interaction (CI) analysis, and normal mode dynamics analysis. Computer programs for implementing these techniques are available ; e. g., Rein, Computer-Assisted Modeling of Receptor-Ligand Interactions (Alan Liss, New York, 1989). Methods for the preparation of chemical derivatives and analogues are well known to those skilled in the art and are described in, for example, Beilstein, Handbook of Organic Chemistry, Springer edition New York Inc., 175 Fifth Avenue, New York, N. Y. 10010 U. S. A. and Organic Synthesis, Wiley, New York, USA. Furthermore, said derivatives and analogues can be tested for their effects according to methods known in the art. Furthermore, peptidomimetics and/or computer aided design of appropriate derivatives and analogues can be used, for example, according to the methods described above.

The inhibitor or antagonist identified by the above-described method are expected prove useful as EBV antiviral agents. The inhibitors and antagonists of the present invention preferably have a specificity at least substantially identical to the binding specificity of the natural ligand or binding partner of the polypeptide described above. An antagonist or inhibitor can have a binding affinity to said polypeptide of at least 105M-1, preferably higher than 107 M-1 and advantageously up to 1010M-1.

In a preferred embodiment, an inhibitor, e. g. suppressive antibody, has an affinity of at least about 10-7 M, preferably at least about 10-9 M and most preferably at least about 10-11 M; and the antagonist has an affinity of less than about 10-7 M, preferably less than about 10-9 M and most preferably in order of 10-"M.

In the case of nucleic acid molecules it is preferred that they have a binding affinity to those encoding the amino acid sequences of a polypeptide selected from the group consisting of TRRAP, ATF5, elF1 and SUPT6H, or a fragment, derivative or ortholog thereof or at most 2-, 5-or 10-fold less than an exact complement of 20 consecutive nucleotides of the above described nucleic acid molecules.

In another embodiment, the present invention relates to a method for producing a therapeutic agent comprising synthesizing the above-described antagonist or inhibitor.

Preferably, the compound identified according to the above described method or its analog or derivative is further formulated in a therapeutical active form or in a form suitable for the application against bacterial infections or diseases related to such an infection. For example, it can be combined with a pharmaceutical acceptable carrier known in the art. Thus, the present invention also relates to a method of producing a (therapeutical effective) composition comprising the steps of one of the above described methods of the invention and combining the compound obtained or identified in the method of the invention or an analog or derivative thereof with a pharmaceutical acceptable carrier.

Also, the present invention relates to a composition comprising the antagonist or inhibitor mentioned above. As is evident from the above, the present invention generally relates to compositions comprising at least one of the aforementioned antagonists or inhibitors, which may be nucleic acid molecules, proteins or antibodies. Advantageously, said composition is for use as a medicament, a diagnostic means, or a kit.

The term"composition", as used in accordance with the present invention, comprises at least one small molecule or-molecule as identified herein above, such as a protein, an antigenic fragment of said protein, a fusion protein, a nucleic acid molecule and/or an antibody as described above and, optionally, further molecules, either alone or in combination, like e. g. molecules which are capable of optimizing antigen processing, cytokines, immunoglobulins, lymphokines or CpG-containing DNA stretches or, optionally, adjuvants. The composition may be in solid, liquid or gaseous form and may be, inter alia, in form of (a) powder (s), (a) tablet (s), (a) solution (s) or (an) aerosol (s). In a preferred embodiment, said composition comprises at least two, preferably three, more preferably four, most preferably five differentially synthesized proteins.

The invention further relates to a method of producing a pharmaceutical composition comprising an antagonist or inhibitor as described above comprising the steps of (a) modifying an inhibitor identified by the method of the invention as a lead compound to achieve (i) modified site of action, spectrum of activity, organ specificity, and/or (ii) improved potency, and/or (iii) decreased toxicity (improved therapeutic index), and/or (iv) decreased side effects, and/or (v) modified onset of therapeutic action, duration of effect, and/or (vi) modified pharmakinetic parameters (resorption, distribution, metabolism and excretion), and/or (vii) modified physico-chemical parameters (solubility, hygroscopicity, color, taste, odor, stability, state), and/or (viii) improved general specificity, organ/tissue specificity, and/or (ix) optimized application form and route by (i) esterification of carboxyl groups, or (ii) esterification of hydroxyl groups with carbon acids, or (iii) esterification of hydroxyl groups to, e. g. phosphates, pyrophosphates or sulfates or hemi succinates, or (iv) formation of pharmaceutically acceptable salts, or (v) formation of pharmaceutically acceptable complexes, or (vi) synthesis of pharmacologically active polymers, or (vii) introduction of hydrophilic moieties, or (viii) introduction/exchange of substituents on aromates or side chains, change of substituent pattern, or (ix) modification by introduction of isosteric or bioisosteric moieties, or (x) synthesis of homologous compounds, or (xi) introduction of branched side chains, or (xii) conversion of alkyl substituents to cyclic analogues, or (xiii) derivatisation of hydroxyl group to ketales, acetales, or (xiv) N-acetylation to amides, phenylcarbamates, or (xv) synthesis of Mannich bases, imines, or (xvi) transformation of ketones or aldehydes to Schiff's bases, oximes, acetales, ketales, enolesters, oxazolidines, thiozolidines or combinations thereof; and (b) formulating the product of said modification with a pharmaceutical acceptable carrier.

The various steps recited above are generally known in the art. They include or rely on quantitative structure-action relationship (QSAR) analyses (Kubinyi, 1993), combinatorial biochemistry, classical chemistry and others (see, for example, Holzgrabe and Bechtold, 2000).

The antagonists and inhibitors of the invention appear to function against gene products which are essential for EBV infection. Accordingly, the above-described antagonists and inhibitors can be used to treat and inhibit EBV infection. Thus, the present invention also relates to a method of producing a therapeutic agent comprising the steps of the methods described hereinbefore and synthesizing the antagonist or inhibitor obtained or identified as described above or an analog or derivative thereof, preferably in an amount sufficient to provide said agent in a therapeutical effective amount to a patient.

Compounds identified by the above methods or analogs are formulated for therapeutic use as pharmaceutical compositions. The compositions can also include, depending on the formulation desired, pharmaceutical acceptable, usually sterile, non-toxic carriers or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological saline, Ringer's solutions, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include other carriers, adjuvants, or nontoxic, nontherapeutic, nonimmunogenic stabilizers and the like.

A therapeutically effective dose refers to that amount of protein or its antibodies, antagonists, or inhibitors which ameliorate the symptoms or condition.

Therapeutic efficacy and toxicity of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e. g., ED50 (the dose therapeutically effective in 50% of the population) and LD50 (the dose lethal to 50% of the population). The dose ratio between therapeutic and toxic effects is the therapeutic index, and it can be expressed as the ratio, LD50/ED50.

Compositions comprising such carriers can be formulated by well known conventional methods. These pharmaceutical compositions can be administered to the subject at a suitable dose. Administration of the suitable compositions may be effected by different ways, e. g., by intravenous, intraperitoneal, subcutaneous, intramuscular, topical, intradermal, intranasal or intrabronchial administration. The dosage regimen will be determined by the attending physician and clinical factors.

As is well known in the medical arts, dosages for any one patient depends upon many factors, including the patient's size, body surface area, age, the particular compound to be administered, sex, time and route of administration, general health, and other drugs being administered concurrently. Proteinaceous pharmaceutically active matter may be present in amounts between 1 ng and 10 mg per dose; however, doses below or above this exemplary range are envisioned, especially considering the aforementioned factors. Administration of the suitable compositions may be effected by different ways, e. g., by intravenous, intraperitoneal, subcutaneous, intramuscular, topical or intradermal administration.

If the regimen is a continuous infusion, it should also be in the range of 1 Ng to 10 mg units per kilogram of body weight per minute, respectively. Progress can be monitored by periodic assessment. The compositions of the invention may be administered locally or systemically. Administration will generally be parenterally, e. g., intravenously. The compositions of the invention may also be administered directly to the target site, e. g., by biolistic delivery to an internal or external target site or by catheter to a site in an artery. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.

Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.

Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, cheating agents, and inert gases and the like. Furthermore, the pharmaceutical composition of the invention may comprise further agents such as interleukins, interferons and/or CpG-containing DNA stretches, depending on the intended use of the pharmaceutical composition.

In another embodiment, the present invention relates to a kit comprising at least one of the aforementioned antagonists or inhibitors of the invention. The kit of the invention as well as the composition may in a preferred embodiment contain further ingredients such as selection markers, antibiotics, cytokines and components for simplifying or supporting the treatment of EBV infection or disorders or diseases related to EBV infections. The kit of the invention may advantageously be used for carrying out the method of the invention and could be, inter alia, employed in a variety of applications referred to herein, e. g., in the diagnostic field or as research tool. The parts of the kit of the invention can be packaged individually in vials or in combination in containers or multicontainer units. Manufacture of the kit follows preferably standard procedures which are known to the person skilled in the art. The kit or its ingredients according to the invention can be used in antibacterial therapies, for example, for any of the above described methods for detecting further inhibitors and antagonists essential for bacterial growth and survival. The kit of the invention and its ingredients are expected to be very useful for the healing and protection of animals and humans suffering from an EBV infection.

The present invention also relates to a method for treating or preventing EBV infections or diseases or disorders related to EBV infections comprising the step of administering to a subject in need thereof an antagonist or inhibitor identified herein above, optionally comprised in a pharmaceutical composition of the invention.

In another embodiment the present invention relates to the use of a polypeptide encoded by the gene as identified above or a fragment, derivative or ortholog thereof or of any of said genes for the identification of an antagonist or inhibitor of said polypeptide fragment, derivate or ortholog or said gene, in particular for the use as an antiviral agent.

In a further embodiment the present invention relates to the use of said polypeptide, the therapeutic agent produced according to the invention, the antagonist or inhibitor obtained or identified by the method or use according to the invention for the preparation of a pharmaceutical composition for the treatment of EBV infection, a disorder and/or disease related to EBV infections.

In another embodiment the present invention relates to a method for treating or preventing EBV infection or diseases or disorders related to EBV infection comprising the step of administering to a subject in need thereof an antagonist or inhibitor identified herein above, optionally comprised in the pharmaceutical composition according to the present invention.

In a preferred embodiment of the above described uses, methods, compositions and kits said antagonist or inhibitor is or is derived from an antisense nucleic acid molecule to the gene encoding said polypeptide, an antibody, an inactive form of said polypeptide, a natural ligand or binding partner of said polypeptide, or from a molecule which captures the natural ligand or binding partner of said polypeptide.

Examples for such antagonists and inhibitors are known to the person skilled in the art and are also described above in context with the information to TRRAP, ATF5, elF1 and SUPT6H.

In another preferred embodiment of the above described uses and methods of the present invention said disease or disorder related to EBV infection is EBV- associated hemophagocytic syndrome, chronic active EBV infection, T-cell lymphoma,-natural killer cell leukemia/lymphoma, lymphoproliferative diseases in immunocompromised hosts, Hodgkin's disease, pyothorax-associated B-cell lymphom, smooth-muscle tumors, gastric carcinoma or Burkitt's lymphom.

In a further embodiment the present invention relates to the use of the above- described polypeptide, a fragment, derivative or ortholog thereof or of any of said genes for screening for polypeptides interacting with said polypeptide using protein-protein interaction technologies, and/or for validating such interaction as being essential for EBV infection and/or for screening for antagonists or inhibitors of such interaction.

In a further embodiment the present invention relates to the use of the above- described polypeptide, a fragment, derivative or ortholog thereof or of any of said genes for screening of polypeptide for polypeptide binding to said polypeptide, and/or for validating the peptides binding to said polypeptide as preventing EBV infection and/or for screening for small molecules competitively displacing said peptides.

In another embodiment the present invention relates to the use of a conditional mutant of a gene as described above or a fragment, derivative or ortholog thereof or of surrogate ligands against said gene expressed in a cell infected by EBV to induce a lethal phenotype in the cell and/or for the analysis of said cell for surrogate markers by comparison of RNA or protein profiles in said cell with RNA or protein profiles in non-infected cells, and/or the use of said surrogate markers for the identification of antagonists of the function of said gene.

In another embodiment the present invention relates to a method for identifying or isolating a surrogate marker comprising the steps as described in the above- recited method of the present invention.

As evident from the example and the description above the findings of the present invention can be generally applied to the identification of genes associated with infection by identifying genes that are regulated in terms of expression in the course of an infection. Preferably, said infection is a viral infection, most preferably an EBV infection. Accordingly, the present invention also relates to a method to identify genes associated with an infection comprising identifying genes that are regulated in terms of expression in the course of an infection, preferably an infection with Epstein-Barr virus. In a preferred embodiment of the method said reference gene is chosen from the list of c-myc, LMP1, EP11, or cdk4.

In another embodiment the present invention relates to a nucleic acid sequence and amino acid sequence identified by a method described above1.

In addition embodiment the present invention relates to a method of identifying an antagonist or inhibitor of infection, said method comprising (a) testing a candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors for the inhibition or reduction of transcription of a gene identified by a method described above, or of a nucleic acid sequence of the invention; or (b) testing a candidate antagonist or inhibitor, or a sample comprising a plurality of said candidate antagonists or inhibitors for the inhibition or reduction of translation of mRNA transcribed from said gene or nucleic acid sequence or a fragment or derivative or ortholog thereof; and (c) determining whether said candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors tests positive in step (a) and/or (b).

In another embodiment the present invention relates to a method for of identifying an antagonist or inhibitor of infection, said method comprising the steps of (a) contacting a polypeptide comprising an amino acid sequence of the invention or a fragment, derivative or ortholog thereof with a candidate antagonist or inhibitor or a sample comprising a plurality of said candidate antagonists or inhibitors; and (b) determining whether said antagonist or inhibitor binds to said polypeptide.

It is to be understood that any one of the above described embodiments particularly and exemplary explained for antagonists and inhibitors of EBV infection can be equally applied to the later embodiments of the present invention.

This also holds true for the embodiments characterized in the claims.

These and other embodiments are disclosed and encompassed by the description and examples of the present invention. Further literature concerning any one of the methods, uses and compounds to be employed in accordance with the present invention may be retrieved from public libraries, using for example electronic devices. For example the public database"Medline"may be utilized which is available on the Internet, for example under http://www. ncbi. nim. nih. gov/PubMed/medline. html. Further databases and addresses, such as http ://www. ncbi. nim. nih. gov/, http://www. infobiogen. fr/, http://www. fmi. ch/biology/research_tools. html, http ://ww w. tigr. org/, are known to the person skilled in the art and can also be obtained using, e. g., http ://www. lycos. com. An overview of patent information in biotechnology and a survey of relevant sources of patent information useful for retrospective searching and for current awareness is given in Berks, TIBTECH 12 (1994), 352-364.

The present invention is further illustrated by reference to the following non- limiting examples.

Unless stated otherwise in the examples, all recombinant DNA techniques are performed according to protocols as described in Sambrook et al. (1989), Molecular Cloning : A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY or in Volumes 1 and 2 of Ausubel et al. (1994), Current Protocols in Molecular Biology, Current Protocols. Standard materials and methods for plant molecular work are described in Plant Molecular Biology Labfase (1993) by R. D. D. Croy, jointly published by BIOS Scientific Publications Ltd. (UK) and Blackwell Scientific Publications (UK).

Example Experiments based on a conditional cellular system for Epstein-Barr virus nuclear antigen 2 (EBNA2) have been performed. In particular, a conditional cellular system for EBNA2 (the ER/EB cell line containing an inducible Estrogenreceptor/EBNA2 fusion protein) has been generated and used for functional studies in EBV immortalized B-cells. In accordance with the present invention expression profiling for a systematic approach towards the identification of novel EBNA2 target genes has been employed for novel EBNA2 target genes.

To this end time course experiments were performed in order to follow the induction of genes post-EBNA2 activation in ER/EB cells. Gene expression analysis ExpressCode (TM) technology has been applied : 9,000 cDNA clones were arrayed at high densities on the 8 x 12 cm nylon membrane and hybridized with 33P-labelled cDNA transcribed from RNA populations (2lig total RNA) of different time points post-stimulation. Statistical confidence of the data was derived by 4 replicates of each experiment. The first goal was to identify genes with an expression profile similar to the c-myc, LMP1, EP11, or cdk4. This was done using the Profile Search function available in the Spotfire. net 5.1 data analysis. By hierarchical clustering of the results, group specific expression profiles for a subset of genes could be identified. Thus, four genes could be identified for the first time to be associated with EBV infection. These are TRRAP, ATF5, elF1 and SUPT6H are further discussed in the description.

Table I TRRAP Amino acid sequence (SEQ ID 1), GenBank accession number NM_003496 <BR> MAFVATQGATVVDQTTLMKKYLQFVAALTDVNTPDETKLKMMQE<BR> VSENFENVTSSPQYSTFLEHIIPRFLTFLQDGEVQFLQEKPAQQLRKLVLEIIHRIPT NEHLRPHTKNVLSVMFRFLETENEENVLICLRIIIELHKQFRPPITQEIHHFLDFVKQ <BR> <BR> IYKELPKWNRYFENPQVIPENTVPPPEMVGMITTIAVKVNPEREDSETRTHSIIPRG< ;BR> SLSLKVLAELPIIWLMYQLYKLNIHNWAEFVPLIMNTIAIQVSAQARQHKLYNKEL YADFIAAQIKTLSFLAYIIRIYQELVTKYSQQMVKGMLQLLSNCPAETAHLRKELLIA AKHILTTELRNQFIPCMDKLFDESILIGSGYTARETLRPLAYSTLADLVHHVRQHLPL&l t;BR> SDLSLAVQLFAKNIDDESLPSSIQTMSCKLLLNLVDCIRSKSEQESGNGRDVLMRMLE&l t;BR> VFVLKFHTIARYQLSAIFKKCKPQSELGAVEAALPGVPTAPAAPGPAPSPAPVPAPPP PPPPPPPATPVTPAPVPPFEKQGEKDKEDKQTFQVTDCRSLVKTLVCGVKTITWGITS CKAPGEAQFIPNKQLQPKETQIYIKLVKYAMQALDIYQVQIAGNGQTYIRVANCQTVR MKEEKEVLEHFAGVFTMMNPLTFKEIFQTTVPYMVERISKNYALQIVANSFLANPTTS&l t;BR> ALFATILVEYLLDRLPEMGSNVELSNLYLKLFKLVFGSVSLFAAENEQMLKPHLHKIV NSSMELAQTAKEPYNYFLLLRALFRSIGGGSHDLLYQEFLPLLPNLLQGLNMLQSGLH KQHMKDLFVELCLTVPVRLSSLLPYLPMLMDPLVSALNGSQTLVSQGLRTLELCVDNL&l t;BR> QPDFLYDHIQPVRAELMQALWRTLRNPADSISHVAYRVLGKFGGSNRKMLKESQKLHY&l t;BR> WTEVQGPSITVEFSDCKASLQLPMEKAIETALDCLKSANTEPYYRRQAWEVIKCFLV AMMSLEDNKHALYQLLAHPNFTEKTIPNVIISHRYKAQDTPARKTFEQALTGAFMSAV IKDLRPSALPFVASLIRHYTMVAVAQQCGPFLLPCYQVGSQPSTAMFHSEENGSKGMD PLVLIDAIAICMAYEEKELCKIGEVALAVIFDVASIILGSKERACQLPLFSYIVERLC ACCYEQAWYAKLGGWSIKFLMERLPLTWVLQNQQTFLKALLFVMMDLTGEVSNGAVA< ;BR> MAKTTLEQLLMRCATPLKDEERAEEIVAAQEKSFHHVTHDLVREVTSPNSTVRKQAMH SLQVLAQVTGKSVTVIMEPHKEVLQDMVPPKKHLLRHQPANAQIGLMEGNTFCTTLQP RLFTMDLNVVEHKVFYTELLNLCEAEDSALTKLPCYKSLPSLVPLRIAALNALAACNY&l t;BR> LPQSREKIIAALFKALNSTNSELQEAGEACMRKFLEGATIEVDQIHTHMRPLLMMLGD&l t;BR> YRSLTLNWNRLTSVTRLFPNSFNDKFCDQMMQHLRKWNEVVVITHKGGQRSDGNEMK< ;BR> ICSAIINLFHLIPAAPQTLVKPLLEVVMKTERAMLIEAGSPFREPLIKFLTRHPSQTV&l t;BR> ELFMMEATLNDPQWSRMFMSFLKHKDARPLRDVLAANPNRFITLLLPGGAQTAVRPGS&l t;BR> PSTSTMRLDLQFQAIKIISIIVKNDDSWLASQHSLVSQLRRVWVSENFQERHRKENMA ATNWKEPKLLAYCLLNYCKRNYGDIELLFQLLRAFTGRFLCNMTFLKEYMEEEIPKNY SIAQKRALFFRFVDFNDPNFGDELKAKVLQHILNPAFLYSFEKGEGEQLLGPPNPEGD&l t;BR> NPESITSVFITKVLDPEKQADMLDSLRIYLLQYATLLVEHAPHHIHDNNKNRNSKLRR&l t;BR> LMTFAWPCLLSKACVDPACKYSGHLLLAHIIAKFAIHKKIVLQVFHSLLKAHAMEARA IVRQAMAILTPAVPARMEDGHQMLTHWTRKIIVEEGHTVPQLVHILHLIVQHFKVYYP VRHHLVQHMVSAMQRLGFTPSVTIEQRRLAVDLSEWIKWELQRIKDQQPDSDMDPNS< ;BR> SGEGVNSVSSSIKRGLSVDSAQEVKRFRTATGAISAVFGRSQSLPGADSLLAKPIDKQ&l t;BR> HTDTVVNFLIRVACQVNDNTNTAGSPGEVLSRRCVNLLKTALRPDMWPKSELKLQWFD&l t;BR> KLLMTVEQPNQVNYGNICTGLEVLSFLLTVLQSPAILSSFKPLQRGIAACMTCGNTKV LRAVHSLLSRLMSIFPTEPSTSSVASKYEELECLYAAVGKVIYEGLTNYEKATNANPS QLFGTLMILKSACSNNPSYIDRLISVFMRSLQKMVREHLNPQAASGSTEATSGTSELV MLSLELVKTRLAVMSMEMRKNFIQAILTSLIEKSPDAKILRAVVKIVEEWVKNNSPMA&l t;BR> ANQTPTLREKSILLVKMMTYIEKRFPEDLELNAQFLDLVNYVYRDETLSGSELTAKLE&l t;BR> PAFLSGLRCAQPLIRAKFFEVFDNSMKRRVYERLLYVTCSQNWEAMGNHFWIKQCIEL&l t;BR> LLAVCEKSTPIGTSCQGAMLPSITNVINLADSHDRAAFAMVTHVKQEPRERENSESKE&l t;BR> EDVEIDIELAPGDQTSTPKTKELSEKDIGNQLHMLTNRHDKFLDTLREVKTGALLSAF&l t;BR> VQLCHISTTLAEKTWVQLFPRLWKILSDRQQHALAGEISPFLCSGSHQVQRDCQPSAL&l t;BR> NCFVEAMSQCVPPIPIRPCVLKYLGKTHNLWFRSTLMLEHQAFEKGLSLQIKPKQTTE&l t;BR> FYEQESITPPQQEILDSLAELYSLLQEEDMWAGLWQKRCKYSETATAIAYEQHGFFEQ&l t;BR> AQESYEKAMDKAKKEHERSNASPAIFPEYQLWEDHWIRCSKELNQWEALTEYGQSKGH INPYLVLECAWRVSNWTAMKEALVQVEVSCPKEMAWKVNMYRGYLAICHPEEQQLSFI ERLVEMASSLAIREWRRLPHVVSHVHTPLLQAAQQIIELQEAAQINAGLQPTNLGRNN&l t;BR> SLHDMKTVVKTWRNRLPIVSDDLSHWSSIFMWRQHHYQAIVTAYENSSQHDPSSNNAM&l t;BR> LGVHASASAIIQYGKIARKQGLVNVALDILSRIHTIPTVPIVDCFQKIRQQVKCYLQL AGVMGKNECMQGLEVIESTNLKYFTKEMTAEFYALKGMFLAQINKSEEANKAFSAAVQ MHDVLVKAWAMWGDYLENIFVKERQLHLGVSAITCYLHACRHQNESKSRKYLAKVLWL&l t;BR> LSFDDDKNTLADAVDKYCIGVPPIQWLAWIPQLLTCLVGSEGKLLLNLISQVGRVYPQ&l t;BR> AVYFPIRTLYLTLKIEQRERYKSDPGPIRATAPMWRCSRIMHMQRELHPTLLSSLEGI&l t;BR> VDQMVWFRENWHEEVLRQLQQGLAKCYSVAFEKSGAVSDAKITPHTLNFVKKLVSTFG VGLENVSNVSTMFSSAASESLARRAQATAQDPVFQKLKGQFTTDFDFSVPGSMKLHNL ISKLKKWIKILEAKTKQLPKFFLIEEKCRFLSNFSAQTAEVEIPGEFLMPKPTHYYIK IARFMPRVEIVQKHNTAARRLYIRGHNGKIYPYLVMNDACLTESRREERVLQLLRLLN&l t;BR> PCLEKRKETTKRHLFFTVPRWAVSPQMRLVEDNPSSLSLVEIYKQRCAKKGIEHDNP< ;BR> ISRYYDRLATVQARGTQASHQVLRDILKEVQSNMVPRSMLKEWALHTFPNATDYWTFR KMFTIQLALIGFAEFVLHLNRLNPEMLQIAQDTGKLNVAYFRFDINDATGDLDANRPV PFRLTPNISEFLTTIGVSGPLTASMIAVARCFAQPNFKVDGILKTVLRDEIIAWHKKT QEDTSSPLSAAGQPENMDSQQLVSLVQKAVTAIMTRLHNLAQFEGGESKVNTLVAAAN SLDNLCRMDPAWHPWL TRRAP Nucleic acid sequence (SEQ ID 2), GenBank accession number NM003496 1 cgcgccgggg cctggtgctc ggtcggcggg tgctgccgct ttaagcgggg gcgggactgc 61 gcgcggccga gcggttgcga cgagggctcg gctgggggtc gccggggtcg cgggccgggc 121 ctgcaggagc cgggccgccg aggtcggggc tggttgaact catggacctg atacttttct 181 cttgagaagc aaaccagccc aaaagaaaaa tggcgtttgt tgcaacacag ggggccacgg 241 tggttgacca gaccactttg atgaaaaagt accttcagtt tgtggcagct ctcacagatg 301 tgaatacacc tgatgaaaca aagttgaaaa tgatgcaaga agttagtgaa aattttgaga 361 atgtcacgtc atctcctcag tattctacat tcctagaaca tatcatccct cgattcctta 421 catttctcca agatggagaa gttcagtttc ttcaggagaa accagcacag caactgcgga 481 agctcgtact tgaaataatt catagaatac caaccaacga acatcttcgt cctcacacaa 541 aaaatgtttt gtctgtgatg tttcgctttt tagagacgga aaatgaagaa aatgttctta 601 tttgtctaag aataattatt gagctacaca aacagttcag gccaccgatc acacaagaaa 661 ttcatcattt tctggatttt gtgaaacaga tttacaagga gcttccaaaa gtagtgaacc 721 gctactttga gaaccctcaa gtgatccccg agaacacagt gcctccccca gaaatggttg 781 gtatgataac aacgattgct gtgaaagtca acccggagcg tgaggacagt gagactcgaa 841 cacattccat cattccgagg ggatcacttt ctctgaaagt gttggcagaa ttgcccatta 901 ttgttgtttt aatgtatcag ctctacaaac tgaacatcca caatgttgtt gctgagtttg 961 tgcccttgat catgaacacc attgccattc aggtgtctgc acaagcgagg caacataagc 1021 tttacaacaa ggagttgtat gctgacttca ttgctgctca gattaaaaca ttgtcatttt 1081 tagcttacat tatcaggatt taccaggagt tggtgactaa gtattctcag cagatggtga 1141 aaggaatgct ccagttactt tcaaattgtc cagcagagac tgcacacctc agaaaggagc 1201 ttctgattgc tgccaaacac atcctcacca cagagctgag aaaccagttc attccttgca 1261 tggacaagct gtttgatgaa tccatactaa ttggctcagg atatactgcc agagagactc 1321 taaggcccct cgcctacagc acgctggccg acctcgtgca ccatgtccgc cagcacctgc 1381 ccctcagcga cctctccctc gccgtccagc tcttcgccaa gaacatcgac gatgagtccc 1441 tgcccagcag catccagacc atgtcctgca agctcctgct gaacctggtg gactgcatcc 1501 gttccaagag cgagcaggag agtggcaatg ggagagacgt cctgatgcgg atgctggagg 1561 ttttcgttct caaattccac acaattgctc ggtaccagct ctctgccatt tttaagaagt 1621 gtaagcctca gtcagaactt ggagccgtgg aagcagctct gcctggggtg cccactgccc 1681 ctgcagctcc tggccctgct ccctccccag cccctgtccc tgccccacct ccacccccgc 1741 ccccaccccc acctgccacc cctgtgaccc cggcccccgt gcctcccttc gagaagcaag 1801 gagaaaagga caaggaagac aagcagacat tccaagtcac agactgtcga agtttggtca 1861 aaaccttggt gtgtggtgtc aagacaatca cgtggggcat aacatcatgc aaagcacctg 1921 gtgaagctca gttcattccc aacaagcagt tacaacccaa agagacacag atttacatca 1981 aacttgtgaa atatgcaatg caagctttag atatttatca ggtccagata gcaggaaatg 2041 gacagacata catccgtgtg gccaactgcc agactgtgag aatgaaagag gagaaggagg 2101 tattggagca tttcgctggt gtgttcacaa tgatgaaccc cttaacgttc aaagaaatct 2161 tccaaactac ggtcccttat atggtggaga gaatctcaaa aaattatgct cttcagattg 2221 ttgccaattc cttcttggca aatcctacta cctctgctct gtttgctacg attctggtgg 2281 aatatctcct tgatcgcctg ccagaaatgg gctccaacgt ggagctctcc aacctgtacc 2341 tcaagctgtt caagctggtc tttggctctg tctccctctt tgcagctgaa aatgaacaaa 2401 tgctgaagcc tcacttgcac aagattgtga acagctctat ggagctcgcg cagactgcca 2461 aggaacccta caactacttc ttgctgctac gggcgctgtt tcgctctatt ggtggaggta 2521 gccacgatct cttgtatcag gagttcttgc ctctccttcc aaacctcctg caagggctga 2581 acatgcttca gagtggcctg cacaagcagc acatgaagga cctctttgtg gagctgtgtc 2641 tcaccgtccc tgtgcggctg agctcgcttt tgccgtacct gcccatgctt atggatccct 2701 tggtgtctgc actcaatggg tctcagacat tggtcagcca aggcctcagg acgctggagc 2761 tgtgtgtgga caacctgcag cccgacttcc tctacgacca catccagccg gtgcgcgcag 2821 agcttatgca ggctctgtgg cgcaccttac gcaaccctgc tgacagcatc tcccacgtgg 2881 cctaccgtgt gctcggtaag tttggcggca gtaacaggaa gatgctgaag gagtcgcaga 2941 agctgcacta cgttgtgacc gaggttcagg gccccagcat cactgtggag ttttccgact 3001 gcaaagcttc tctccagctc cccatggaga aggccattga aactgctctg gactgcctga 3061 aaagcgccaa cactgagccc tactaccgga ggcaggcgtg ggaagtgatc aaatgcttcc 3121 tggtggccat gatgagcctg gaggacaaca agcacgcact ctaccagctc ctggcacacc 3181 ccaactttac agaaaagacc atccccaatg ttatcatctc acatcgctac aaagcccagg 3241 acactccagc ccggaagact tttgagcagg ccctgacagg cgccttcatg tctgctgtca 3301 ttaaggacct gcggcccagc gccctgccct ttgtcgccag cttgatccgc cactatacga 3361 tggtggcagt cgcccagcag tgtggccctt tcttgctgcc ttgctaccag gtgggcagcc 3421 agcccagcac agccatgttt cacagtgaag aaaatggctc gaaaggaatg gatcctttgg 3481 ttctcattga tgcaattgct atttgtatgg catatgaaga aaaggagctt tgcaaaatcg 3541 gggaggtggc cctagctgtg atatttgatg ttgcaagtat catcctgggc tccaaggaga 3601 gggcctgcca gctgcccctg ttttcttaca tcgtggagcg cctgtgtgca tgttgttatg 3661 aacaggcgtg gtatgcaaag ctggggggtg tggtgtctat taagtttctc atggagcggc 3721 tgcctctcac ttgggttctc cagaaccagc agacattcct gaaagcactt ctctttgtca 3781 tgatggactt aactggagag gtttccaatg gggcagtcgc tatggcaaag accacgctgg 3841 agcagcttct gatgcggtgc gcaacgcctt taaaagacga ggagagagcc gaagagatcg 3901 tggccgccca ggaaaagtct ttccaccatg tgacacacga cttggttcga gaagtcacct 3961 ctccaaactc cactgtgagg aagcaggcca tgcattcgct gcaggtgttg gcccaggtca 4021 ctgggaagag tgtcacggtg atcatggaac cccacaaaga ggtcctgcag gatatggtcc 4081 cccctaagaa gcacctgctc cgacaccagc ctgccaacgc acagattggc ctgatggagg 4141 ggaacacgtt ctgtaccacg ttgcagccca ggctcttcac aatggacctt aacgtggtgg 4201 agcataaggt gttctacaca gagctgttga atttgtgtga ggctgaagat tcagctttaa 4261 caaagctgcc ctgttataaa agccttccgt cactcgtacc tttacgaatt gcggcattaa 4321 atgcacttgc tgcctgcaat taccttcctc agtccaggga gaaaatcatc gctgcactct 4381 tcaaagccct gaattccacc aatagtgagc tccaagaggc cggagaagcc tgtatgagaa 4441 agtttttaga aggtgctacc atagaagtcg atcaaatcca cacacatatg cgacctttgc 4501 tgatgatgct gggagattac cggagcttga cgctgaatgt tgtgaatcgc ctgacttcgg 4561 tcacgaggct cttcccaaat tccttcaatg ataaattttg tgatcagatg atgcaacatc 4621 tgcgcaagtg gatggaagtg gtggtgatca cccacaaagg gggccagagg agcgacggaa 4681 acgaaatgaa gatttgctca gcaattataa acctttttca tctgatcccg gctgctcctc 4741 agacactggt gaagcctttg ctagaggttg tcatgaaaac ggagcgggcg atgctgatcg 4801 aggcggggag tccattccga gagcccctga tcaagttcct gactcgacat ccctcgcaga 4861 cagtggagct gttcatgatg gaagccacac tgaacgatcc ccagtggagc agaatgttta 4921 tgagtttttt aaaacacaaa gacgccagac ctctgcggga tgtgctggct gccaacccca 4981 acaggttcat caccctgctg ctgccggggg gtgcccagac ggctgtgcgc cccggttcgc 5041 ccagcaccag caccatgcgc ctggacctcc agttccaggc catcaagatc ataagcatta 5101 tagtgaaaaa cgatgactcc tggctggcca gccagcactc tctggtgagc cagttgcgac 5161 gtgtgtgggt gagtgagaac ttccaagaga ggcaccgcaa ggagaacatg gcagccacca 5221 actggaagga gcccaagctg ctggcctact gcctgctgaa ctactgcaaa aggaattacg 5281 gagatataga attgctgttc cagctgctcc gagcctttac tggtcgtttt ctctgcaaca 5341 tgacattctt aaaagagtat atggaggaag agattcccaa aaattacagc atcgctcaga 5401 aacgtgccct gttctttcgc tttgtagact tcaacgaccc caacttcgga gatgaattaa 5461 aagctaaagt tctgcagcat atcttgaatc ctgctttctt gtacagcttt gagaaggggg 5521 aaggagagca gctcttggga cctcccaatc cagaaggaga taacccagaa agcatcacca 5581 gtgtgtttat taccaaggtc ctggaccccg agaagcaggc ggacatgctg gactcgctgc 5641 ggatctacct gctgcagtac gccacgctgc tggtggagca cgccccccac cacatccatg 5701 acaacaacaa gaaccgcaac agcaagctgc gccgcctcat gaccttcgcc tggccctgcc 5761 tgctctccaa ggcctgcgtg gacccagcct gcaagtacag cggacacttg ctcctggcgc 5821 acattatcgc caaattcgcc atacacaaga agatcgtcct gcaggttttt catagtctcc 5881 tcaaggctca cgcaatggaa gctcgagcga tcgtcagaca ggcgatggcc attctgaccc 5941 cggcggtgcc ggccaggatg gaggacgggc accagatgct gacccactgg acccggaaga 6001 tcattgtgga ggaggggcac accgtcccgc agctggtcca cattctgcac ctgatagtgc 6061 aacacttcaa ggtgtactac ccggtacggc accacttggt gcagcacatg gtgagcgcca 6121 tgcagaggct gggcttcacg cccagtgtca ccatcgagca gaggcggctg gccgtggacc 6181 tgtctgaagt cgtcatcaag tgggagctgc agaggatcaa ggaccagcag ccggattcag 6241 atatggaccc aaattccagt ggagaaggag tcaattctgt ctcatcctcc attaagagag 6301 gcctgtccgt ggattctgcc caggaagtga aacgctttag gacggccacc ggagccatca 6361 gtgcagtctt tgggaggagc cagtcgctac ctggagcaga ctctctcctc gccaagccca 6421 ttgacaagca gcacacagac actgtggtga acttccttat ccgcgtggcc tgtcaggtta 6481 atgacaacac caacacagcg gggtcccctg gggaggtgct ctctcgccgg tgtgtgaacc 6541 ttctgaagac tgcgttgcgg ccagacatgt ggcccaagtc cgaactcaag ctgcagtggt 6601 tcgacaagct gctgatgact gtggagcagc caaaccaagt gaactatggg aatatctgca 6661 cgggcctaga agtgctgagc ttcctgctaa ctgtcctcca gtccccagcc atcctcagta 6721 gcttcaaacc tctgcagcgt ggaattgccg cctgcatgac atgtggaaac accaaggtgt 6781 tgcgagccgt ccacagcctt ctctcgcgcc tgatgagcat tttcccaaca gagccgagta 6841 cttccagtgt ggcctccaaa tatgaagagc tggagtgcct ctacgcagcc gtcggaaagg 6901 tcatctatga agggctcacc aactacgaga aggccaccaa tgccaatccc tcccagctct 6961 tcgggaccct tatgatcctc aagtctgcct gcagcaacaa ccccagctac atagacaggc 7021 tgatctccgt ctttatgcgc tccctgcaga agatggtccg ggagcattta aaccctcagg 7081 cagcgtcagg aagcaccgaa gccacctcag gtacaagcga gctggtgatg ctgagtctgg 7141 agctggtgaa gacgcgcctg gcagtgatga gcatggagat gcggaagaac ttcatccagg 7201 ccatcctgac atccctcatc gaaaaatcac cagatgccaa aatcctccgg gctgtggtca 7261 aaatcgtgga agaatgggtc aagaataact ccccaatggc agccaatcag acacctacac 7321 tccgggagaa gtccattttg cttgtgaaga tgatgactta catagaaaaa cgctttccgg 7381 aagaccttga attaaatgcc cagtttttag atcttgttaa ctatgtctac agggatgaga 7441 ccctctctgg cagcgagctg acggcgaaac ttgagcctgc ctttctctct gggctgcgct 7501 gtgcccagcc actcatcagg gcaaagtttt tcgaggtttt tgacaactcc atgaaacgtc 7561 gtgtctacga gcgcttgctc tatgtgacct gttcgcagaa ctgggaagcc atggggaacc 7621 acttctggat caagcagtgc attgagctgc ttctggccgt gtgtgagaag agcaccccca 7681 ttggcaccag ctgccaagga gccatgctcc cgtccatcac caacgtcatc aacctggccg 7741 atagccacga ccgtgccgcc ttcgccatgg tcacacatgt caagcaggag ccccgggagc 7801 gggagaacag cgagtccaaa gaggaggatg tagagataga catcgaacta gctcctgggg 7861 atcagaccag cacgcccaaa accaaagaac tttcagaaaa ggacattgga aaccagctgc 7921 acatgctaac caacaggcac gacaagtttc tggacactct ccgagaggtg aagactggag 7981 cgctgctcag cgctttcgtt cagctgtgcc acatttccac gacgctggca gagaagacgt 8041 gggtccagct tttccccaga ttgtggaaga tcctctctga cagacagcag catgcactcg 8101 cgggtgagat aagtccattt ctgtgcagcg gcagtcacca ggtgcagcgg gactgccagc 8161 ccagcgcgct gaactgcttt gtggaagcca tgtcccagtg cgtgccgcca atccccatcc 8221 gaccctgcgt cctgaagtac ctggggaaga cacacaacct ctggttccgg tccacgctga 8281 tgttggagca ccaggctttt gaaaagggtc tgagtcttca gattaagccg aagcaaacaa 8341 cggagtttta tgagcaggag agcatcaccc cgccgcagca ggagatactg gattcccttg 8401 cggagcttta ctccctgtta caagaggaag atatgtgggc tggtctgtgg cagaagcggt 8461 gcaagtactc ggagacagcg actgcgattg cttacgagca gcacgggttc tttgagcagg 8521 cacaagaatc ctatgaaaag gcaatggata aagccaaaaa agaacatgag aggagtaacg 8581 cctcccctgc tattttccct gaataccagc tctgggaaga ccactggatt cgatgctcca 8641 aggaattgaa ccagtgggaa gccctgacgg agtacggtca gtccaaaggc cacatcaacc 8701 cctacctcgt cctggagtgc gcctggcggg tgtccaactg gactgccatg aaggaggcgc 8761 tggtgcaggt ggaagtgagc tgtccgaagg agatggcctg gaaggtgaac atgtaccgcg 8821 gatacctggc catctgccac cccgaggagc agcagctcag cttcatcgag cgcctggtgg 8881 agatggccag cagcctggcc atccgcgagt ggcggcggct gccccacgta gtgtcccacg 8941 tgcacacgcc tctcctacag gcagcccagc aaatcatcga actccaggaa gctgcacaaa 9001 tcaacgcagg cttacagcca accaacctgg gaaggaacaa cagcctgcac gacatgaaga 9061 cggtggtgaa gacctggagg aaccgactgc ccatcgtgtc tgacgacttg tcccactgga 9121 gcagcatctt catgtggagg cagcatcatt accaggcgat tgtaactgcc tatgagaata 9181 gctctcagca tgatcccagt tcaaataacg ctatgcttgg ggttcatgca tcagcttcag 9241 cgatcatcca gtatggaaaa atcgcccgga aacaaggact ggtcaatgta gctctggata 9301 tattaagtcg gattcatact attccaactg ttcctatcgt ggattgcttc cagaagattc 9361 gacagcaagt taaatgctac ctccagctgg caggcgtcat gggcaaaaac gagtgcatgc 9421 agggccttga agttattgaa tctacaaatt taaaatactt cacaaaagag atgacagccg 9481 aattttatgc actgaaggga atgttcttgg ctcagatcaa caagtccgag gaggcaaaca 9541 aagccttctc tgcagctgtg cagatgcacg atgtgctggt gaaagcctgg gccatgtggg 9601 gcgactacct ggagaacatc tttgtgaagg agcggcagct gcacctgggc gtgtctgcca 9661 tcacctgcta cctgcacgcc tgccggcatc agaacgagag caaatcgagg aaatacttag 9721 ccaaggtgct gtggcttttg agttttgatg atgacaaaaa cactttggca gatgccgtcg 9781 acaagtactg cattggtgtg ccacccatcc agtggctggc ctggatccca cagctgctca 9841 cctgcctggt tggctcggag ggaaagctgc tcttgaacct cattagccag gttggacgcg 9901 tgtatcccca agcggtctac tttcccatcc ggaccctgta cctgaccctg aaaatagaac 9961 agcgggaacg ctacaagagc gatccagggc ccataagagc aacagcaccc atgtggcgct 10021 gcagccgaat catgcacatg cagcgagagc tccaccccac ccttctgtct tccctggaag 10081 gcatcgtcga tcagatggtc tggttcagag aaaattggca tgaagaggtt ctcaggcagc 10141 tccaacaggg cctggcgaaa tgttactccg tggcgtttga gaaaagtgga gcggtgtccg 10201 atgctaaaat caccccccac actctcaatt ttgtgaagaa gttggtgagc acgtttgggg 10261 tgggcctgga gaatgtgtcc aacgtctcga ccatgttctc cagcgcagcc tctgagtctc 10321 tggcccggcg ggcgcaggcc actgcacaag accctgtctt tcagaagctg aaaggccagt 10381 tcacgacgga ttttgacttc agcgttccag gatccatgaa gcttcataat cttatttcta 10441 agttgaaaaa gtggatcaaa atcttggagg ccaagaccaa gcaactcccc aaattcttcc 10501 tcatagagga aaagtgccgg ttcttgagca atttctcggc acagacagct gaagtggaaa 10561 ttcctgggga gtttctgatg ccaaagccaa cgcattatta catcaagatt gcacggttca 10621 tgccccgggt agagattgtg cagaagcaca acaccgcagc ccggcggctg tacatccggg 10681 gacacaatgg caagatctac ccatacctcg tcatgaacga cgcctgcctc acagagtcac 10741 ggcgagagga gcgtgtgttg cagctgctgc gtctgctgaa cccctgtttg gagaagagaa 10801 aggagaccac caagaggcac ttgtttttca cagtgccccg ggttgtggca gtttccccac 10861 agatgcgcct cgtggaggac aacccctctt cactttccct tgtggagatc tacaagcagc 10921 gctgcgccaa gaagggcatc gagcatgaca accccatctc ccgttactat gaccggctgg 10981 ctacggtgca ggcgcgggga acccaagcca gccaccaggt cctccgcgac atcctcaagg 11041 aggttcagag taacatggtg ccgcgcagca tgctcaagga gtgggcgctg cacaccttcc 11101 ccaatgccac ggactactgg acgttccgga agatgttcac catccagctg gctctgatag 11161 gcttcgcgga attcgtcctg catttaaata gactcaaccc cgagatgtta cagatcgctc 11221 aggacactgg caaactgaat gttgcctact ttcgatttga cataaacgac gcgactggag 11281 acctggatgc caaccgtcct gtcccatttc gactcacgcc caacatttct gagtttctga 11341 ccaccatcgg ggtctccggc ccgttgacag cgtccatgat tgcggtcgcc cggtgcttcg 11401 cccagccaaa ctttaaggtg gatggcattc tgaaaacggt tctccgggac gagatcattg 11461 cttggcacaa aaaaacacaa gaggacacgt cctctcctct ctcggccgcc gggcagccag 11521 agaacatgga cagccagcaa ctggtgtccc tggttcagaa agccgtcacc gccatcatga 11581 cccgcctgca caacctcgcc cagttcgaag gcggggaaag caaggtgaac accctggtgg 11641 ccgcggcaaa cagcctggac aatctgtgcc gcatggaccc cgcctggcac ccctggctgt 11701 gactgtggcc gccacggcac gcgggaatgt gaagggcgct ccgggctctg agcccgcagc 11761 ttttacgact tctccctgcg tcgttcctta tattcacaga agccccatag tttcactggg 11821 ttgcggttat tttcctggta gtttgcgtgt aagaaaggga gaatatagtt ttagaggaag 11881 ctgaactatg acgatgctgg gcgaacggtt tgggaaatgg cagagctgaa acttattcca 11941 agctttcaaa ataatctttt aagaagccag gattctccgg tctggaattt ctgagtgagt 12001 ccttttttta tggtgtcctc cctctgtgaa tgtacaggcg gaactgtacg aacagctccc 12061 ttccatccat ttttaactct ttcggaaata acacctcaca gcagcttcgt gcttttgtac 12121 agacctttgt aacaagtgta cagaaaactc attttgtttg agaaacagga gttgatgaac 12181 ccatcatgct ggtttttctc tgagcacaaa gttttaggct gtacacagcc agccttggga 12241 atctcgttga gcgttcggcg tggatccacg gggccaggcc accctgcggg aggccacacg 12301 catccacttc ggattcagtg ggtgaagaca gaactctgag agtctgcagg cggctcctgt 12361 gctttttatt tctggctctt cggatgtctt ctagacattt actatcactg cacctgaaga 12421 aaaaatcact tttaccttcc taatttaaaa agacaaaaca gaaatgtacg ttccttcgct 12481 agctttagtc tttctgttcc catttttata aatctgagca ttgataatgt tctatctaaa 12541 tttgtacagt gtgatttttt ttt ATF5 Amino acid sequence (SEQ ID 3), GenBank accession number NM_012068 MSLLATLGLELDRALLPASGLGWLVDYGKLPPAPAPLAPYEVLG GALEGGLPVGGEPLAGDGFSDWMTERVDFTALLPLEPPLPPGTLPQPSPTPPDLEAMA SLLKKELEQMEDFFLDAPPLPPPSPPPLPPPPLPPAPSLPLSLPSFDLPQPPVLDTLD LLAIYCRNEAGQEEVGMPPLPPPQQPPPPSPPQPSRLAPYPHPATTRGDRKQKKRDQN KSAALRYRQRKRAEGEALEGECQGLEARNRELKERAESVEREIQYVKDLLIEVYKARS QRTRSC ATF5 Nucleic acid sequence (SEQ ID 4), GenBank accession number NM_012068 1 attcattccc tgtcctcgga tcacagtctc ttctcactac agtgtcgccg cctctgcctg 61 cgtagccccg gccatggctc tgtagcctcg acccctttgt gcccccggcc cgtctccgcg 121 ctcaccacgc ctgcgctctc cgctcccacc ttctttcttc agccgaggcc gccgccgcct 181 ctccttgctg cagccatgga gtcttccact ttcgccttgg tgcctgtctt cgcccacctg 241 agcatcctcc agagcctcgt gccagctgct ggtgcagcct ctcctgttgc catcagtgcc 301 cagcacctgt gctacagcca tgtcactcct ggcgaccctg gggctggagc tggacagggc 361 cctgctccca gctagtgggc tgggatggct cgtagactat gggaaactcc ccccggcccc 421 tgcccccctg gctccctatg aggtccttgg gggagccctg gagggcgggc ttccagtggg 481 gggagagccc ctggcaggtg atggcttctc tgactggatg actgagcgag ttgatttcac 541 agctctcctc cctctggagc ctcccctacc ccccggcacc ctcccccaac cttccccaac 601 cccacctgac ctggaagcta tggcctccct cctcaagaag gagctggaac agatggaaga 661 cttcttccta gatgccccgc ccctcccacc accctccccg ccgccactac caccaccacc 721 actaccacca gccccctccc tccccctgtc cctcccctcc tttgacctcc cccagccccc 781 tgtcttggat actctggact tgctggccat ctactgccgc aacgaggccg ggcaggagga 841 agtggggatg ccgcctctgc ccccgccaca gcagccccct cctccttctc cacctcaacc 901 ttctcgcctg gccccctacc cacatcctgc caccacccga ggggaccgca agcaaaagaa 961 gagagaccag aacaagtcgg cggctctgag gtaccgccag cggaagcggg cagagggtga 1021 ggccctggag ggcgagtgcc aggggctgga ggcacggaat cgcgagctga aggaacgggc 1081 agagtccgtg gagcgcgaga tccagtacgt caaggacctg ctcatcgagg tttacaaggc 1141 ccggagccag aggacccgta gctgctagaa gggcaggggt gtggcttctg ggggctggtc 1201 ttcagctctg gcgccttcat ccccctgcct ctaccttcat tccaaacccc tctcggccgg 1261 gtgcagtggc ttatgcttgt aatcccagca ctttgggagg ccaaggcagg aggatcgttt 1321 gaggccagga ggtcaatacc agcctgggca acatagtaag accctgtctc tattaaaaaa 1381 aaaaaatcaa cccttcttcc ccaccaaacc acccaactcc tctctactct tatcctttta 1441 tcctctgtct ctgcttatca cctctcttgc gtatttctgg atctccttcc ctcctttctc 1501 gtccaaatca tgaaatgttt ggccttagtc aatgtctatg cccgtcacat aacagccgag 1561 gcaccgaggc ccacagggaa gcagctggga gcttggaaac ctggtctctt gaatttcaaa 1621 cctggtttct tacaggtggt tgtctggggt gggtggagtg gcgacaggat agagctgaag 1681 gactatgcaa atgaggaagt aagtcagggc gggctttgag aaggggaccc atatcctaca 1741 ggcaaaaagc aggctaggtg accttgggac actacgctaa gggagggagg ctaaaggcgg 1801 ccaggtttgc agtgcgggaa gatgagcagg ccagtgggag gaggggcagg gcagggctgt 1861 agttggtgac tgggtgttca ttttagctct aagaaaaaaa atcagtgttt cgtgaaggtg 1921 ttggagaggg gctgtgtctg ggtgagggat ggcggggtac tgattttttt gggaggttat 1981 gagcaaaaat aaaacgaaac atttcctctg eIF1 (A121/SUI1) Amino acid sequence (SEQ ID 5), GenBank accession number NM005801 MSAIQNLHSFDPFADASKGDDLLPAGTEDYIHIRIQQRNGRKTL TTVQGIADDYDKKKLVKAFKKKFACNGTVIEHPEYGEVIQLQGDQRKNICQFLVEIGL&l t;BR> AKDDQLKVHGF eIF1 (A121/SUI1) Nucleic acid sequence (SEQ ID 6), GenBank accession number Nom005801 1 gccgccgycg aggattcagc agcctccccc ttgagccccc tcgcttcccg acgttccgtt 61 cccccctgcc cgccttctcc cgccaccgcc gccgccgcct tccgcagccg tttccaccga 121 ggaaaaggaa tcgtatcgta tgtccgctat ccagaacctc cactctttcg acccctttgc 181 tgatgcaagt aagggtgatg acctgcttcc tgctggcact gaggattata tccatataag 241 aattcaacag agaaacggca ggaagaccct tactactgtc caagggatcg ctgatgatta 301 cgataaaaag aaactagtga aggcgtttaa gaaaaagttt gcctgcaatg gtactgtaat 361 tgagcatccg gaatatggag aagtaattca gctacagggt gaccaacgca agaacatatg 421 ccagttcctc gtagagattg gactggctaa ggacgatcag ctgaaggttc atgggtttta 481 agtgcttgtg gctcactgaa gcttaagtga ggatttcctt gcaatgagta gaatttccct 541 tctctccctt gtcacaggtt taaaaacctc acagcttgta taatgtaacc atttggggtc 601 cgcttttaac ttggactagt gtaactcctt catgcaataa actgaaaaga gccatgcaaa SUPT6H Amino acid sequence (SEQ ID 7), GenBank accession number U46691 MSDDEDDDEEEYGKEEHEKEAIAEEIFQDGEGEEGQEAMEAPMA PPEEEEEDDEESDIDDFIVDDDGQPLKKPKWRKKLPGYTDAALQEAQEIFGVDFDYDE FEKYNEYDEELEEEYEYEDDEAEGEIRVRPKKTTKKRVSRRSIFEMYEPSELESSHLT DQDNEIRATDLPERFQLRSIPVKGAEDDELEEEADWIYRNAFATPTISLQESCDYLDR GQPASSFSRKGPSTIQKIKEALGFMRNQHFEVPFIAFYRKEYVEPELHINDLWRVWQW DEKWTQLRIRKENLTRLFEKMQAYQYEQISADPDKPLADGIRALDTTDMERLKDVQSM DELKDVYNHFLLYYGRDIPKMQNAAKASRKKLKRVREEGDEEGEGDEAEDEEQRGPEL KQASRRDMYTICQSAGLDGLAKKFGLTPEQFGENLRDSYQRHETEQFPAEPLELAKDY VCSQFPTPEAVLEGARYMVALQIAREPLVRQVLRQTFQERAKLNITPTKKGRKDVDEA HYAYSFKYLKNKPVKELRDDQFLKICLAEDEGLLTTDISIDLKGVEGYGNDQTYFEEI KQFYYRDEFSHQVQEWNRQRTMAIERALQQFLYVQMAKELKNKLLAEAKEYVIKACSR <BR> <BR> KLYNWLRVAPYRPDQQVEEDDDFMDENQGKGIRVLGIAFSSARDHPVFCALVNGEGEV&l t;BR> TDFLRLPHFTKRRTAWREEEREKKAQDIETLKKFLLNKKPHVVTVAGENRDAQMLIED&l t;BR> VKRIVHELDQGQQLSSIGVELVDNELAILYMNSKKSEAEFRDYPPVLRQAVSLARRIQ&l t;BR> DPLIEFAQVCSSDEDILCLKFHPLQEHWKEELLNALYCEFINRVNEVGVDVNRAIAH PYSQALIQYVCGLGPRKGTHLLKILKQNNTRLESRTQLVTMCHMGPKVFMNCAGFLKI DTASLGDSTDSYIEVLDGSRVHPETYEWARKMAVDALEYDESAEDANPAGALEEILEN PERLKDLDLDAFAEELERQGYGDKHITLYDIRAELSCRYKDLRTAYRSPNTEEIFNML <BR> <BR> TKETPETFYIGKLIICNVTGIAHRRPQGESYDQAIRNDETGLWQCPFCQQDNFPELSE&l t;BR> VWNHFDSGSCPGQAIGVKTRLDNGVTGFIPTKFLSDKWKRPEERVKVGMTVHCRIMK IDIEKFSADLTCRTSDLMDRNNEWKLPKDTYYDFDAEAADHKQEEDMKRKQQRTTYIK <BR> <BR> RVIAHPSFHNINFKQAEKMMETMDQGDVIIRPSSKGENHLTVTWKVSDGIYQHVDVRE&l t;BR> EGKENAFSLGATLWINSEEFEDLDEIVARYVQPMASFARDLLNHKYYQDCSGGDRKKL EELLIKTKKEKPTFIPYFICACKELPGKFLLGYQPRGKPRIEYVTVTPEGFRYRGQIF PTVNGLFRWFKDHYQDPVPGITPSSSSRTRTPASINATPANINLADLTRAVNALPQNM TSQMFSAIAAVTGQGQNPNATPAQWASSQYGYGGSGGGSSAYHVFPTPAQQPVATPLM TPSYSYTTPSQPITTPQYHQLQASTTPQSAQAQPQPSSSSRQRQQQPKSNSHAAIDWG KMAEQWLQEKEAERRKQKQRLTPRPSPSPMIESTPMSIAGDATPLLDEMDR SUPT6H Nucleic acid sequence (SEQ ID 8), GenBank accession number U46691 1 tgcttgcact cattttctcc actgccaccc tgtgaagagt tgccttccgc catgattgta 61 agtttcccga ggtctcccca gccatgcaga actatgagtc aatcaaatcc cctttctcca 121 cgaataaccc cgtcttgggt atttcttcat agcagcatag gaatggacca acacacttcc 181 tgaatcagaa actcagggga tgaggcccat caatctgtgt ttcaacaaga cttccaggga 241 ctgcgatgtg cactcaagtc tgagaactac accagtaagt gataataaat atgacaacat 301 atttttttga atgctttaat gaaccttatc tattttcctt gcaatgccct gggcttcctc 361 caaatgagta aagaatttct gcaaagtcac ttagggttca gcagaatccc aggtgtggcc 421 aaagaatgtg catgggcagg tagaccatca gtaggttaga actcaagtcc catgatggct 481 ttgaaaaaaa attgatgttg cttgatctga tctcagcttc cagggcacct cttcagggaa 541 ggcttccctg agcttcctgg gctaaggtag aagctcctgt cacattttcc cagaagcatt 601 ccaaatttct gcttcatcat cacagtgctc atcatactgc aaaaggtcta tctgtttaat 661 catctgcttc cttcacccta agcttttttt tttttttttt tttttttttt gagacggagt 721 ctcgctctgt tatccaggct ggagtgcagt ggctccatct ccgctcactg caagctccgc 781 ctccttggtt catgccattc tcctgcctca gcctcctgag tagctgggac tacagccacc 841 cgccaccaca cccggataat tttttgtatt tttagtagag acggggtttc accctgttag 901 ccaggatggt ctcgatctcc tgacctagtg atcctcccgc ctcagactcc caaagtgttg 961 ggattacagg catgagccac tgcgcctggc caccctaagc ttagtgaggg cagagactct 1021 ggtgattcag aagatgatgt tggccacaag aagagaaaac gcacctcttt tgatgaccgc 1081 ctggaggatg atgattttga cctcattgag gagaatttgg gtgtcaaagt caaaagagga 1141 caaaagtacc ggcgtgtcaa aaaaatgtca gatgacgagg acgatgacga ggaggaatat 1201 ggcaaggagg aacatgaaaa agaagctatt gcggaagaaa tcttccagga tggggaaggg 1261 gaagaagggc aggaggccat ggaggccccc atggctcctc cagaggagga ggaagaagat 1321 gatgaggagt cagatattga cgacttcatt gtggatgatg atggacagcc tctgaaaaaa 1381 cctaagtggc ggaaaaagct tcctggatac acagacgcgg ccctgcaaga agcccaggaa 1441 atcttcggtg tggactttga ctatgatgaa tttgagaaat acaatgagta tgatgaagaa 1501 ctggaggaag agtatgagta tgaggatgat gaggctgagg gtgaaatccg agtgcgcccc 1561 aagaagacca ccaagaagcg tgtgagccgt aggagcatct ttgaaatgta tgagcccagt 1621 gagctagaaa gcagccacct cacagatcag gacaatgaaa tccgagccac tgacctgcct 1681 gagaggttcc agctccgctc catcccagtc aagggggctg aagatgatga actagaagaa 1741 gaagctgact ggatctacag gaatgctttt gccacaccaa ccatttctct ccaggaaagc 1801 tgtgattacc tagaccgagg gcagccagcc agcagcttca gtcggaaagg gcccagcaca 1861 attcagaaga tcaaagaggc cctgggcttc atgcgaaatc agcattttga ggtgcctttt 1921 attgccttct atcgaaagga gtatgtggag cctgagttgc acatcaatga cctatggaga 1981 gtctggcagt gggatgaaaa gtggacccag ctgcggatcc gtaaagagaa cctaacacgg 2041 ctgtttgaga agatgcaggc ttatcagtat gaacagatct ctgctgaccc tgacaaacct 2101 cttgctgatg gcatccgggc tctggacacc actgacatgg agaggctcaa ggatgtccaa 2161 tcaatggatg agctgaaaga tgtctacaac cattttcttc tttattatgg ccgagacatc 2221 cctaagatgc agaacgccgc caaagctagc cgcaagaagc tgaagcgtgt cagggaagag 2281 ggagatgaag aaggtgaagg tgacgaggca gaagatgagg agcagagggg gcctgagctc 2341 aagcaagcct ctcgccgaga catgtacacc atctgccaga gtgctgggct agatggtctg 2401 gccaaaaagt ttgggcttac tcccgagcag tttggggaga acctgcggga tagctaccag 2461 cggcacgaga cagagcagtt tcccgcggag cccttggagc tggccaagga ttacgtttgc 2521 agccagttcc ctactccaga agctgtgcta gaaggcgccc gctacatggt agccctgcag 2581 attgcccgtg agccccttgt ccggcaggtg ctgaggcaaa ccttccaaga gagagccaag 2641 ttaaatataa cccccaccaa gaaaggtaga aaggatgtgg atgaggccca ctatgcctat 2701 tccttcaagt atttaaagaa caagcctgtt aaggaactga gagatgacca gtttctcaag 2761 atatgcctgg ctgaagacga agggctcctc accactgaca tcagcataga tttgaaggga 2821 gtggaaggct atggcaacga ccagacatat tttgaggaga taaaacagtt ttactaccga 2881 gatgagttca gccaccaggt gcaggagtgg aaccggcagc gcaccatggc catcgaacgg 2941 gctttacagc agttcctcta tgtgcagatg gccaaagaac tcaagaacaa gctgctggct 3001 gaagccaagg aatatgtcat aaaggcctgt agtcgaaagc tctacaattg gttgagagtg 3061 gcaccctacc gaccagatca gcaggtggaa gaagatgacg actttatgga cgagaaccaa 3121 gggaagggca ttcgagtcct cggcattgct ttctcctctg ccagagatca ccctgtgttc 3181 tgcgccctgg tcaatggtga aggagaagtg acagacttcc ttcgactgcc ccattttacc 3241 aaacggcgaa ctgcatggag agaggaagag cgggaaaaga aggctcaaga cattgaaacg 3301 ctaaagaaat ttctcctgaa taagaagcct catgtagtga cagttgcagg agagaacagg 3361 gacgcccaga tgttgattga agatgtgaag cgcattgtac atgagctgga ccagggccag 3421 cagctgtcat ctattggggt agagctggtt gacaacgagt tggccattct ctatatgaac 3481 agcaagaagt cagaggcaga gttccgggat tatcctccag tgctgagaca ggccgtctcc 3541 ctggcccggc gcatccagga ccctctgatt gaatttgccc aggtgtgcag ttccgatgaa 3601 gacatcctgt gtctcaagtt tcaccccttg caggagcatg tggtgaaaga ggagctgctc 3661 aacgccttgt actgtgaatt tatcaaccga gtcaatgagg tcggggtcga tgtcaaccgt 3721 gccattgccc acccttacag ccaggccttg atccagtatg tttgtggcct gggacctcgg 3781 aaagggaccc acctcctgaa gatcctgaag cagaacaaca cccggctcga gagccggacc 3841 cagctggtca ccatgtgcca catgggtccc aaagtcttca tgaattgtgc tggcttcctc 3901 aagatcgaca cggcctccct gggggacagc actgactcat atattgaagt ccttgatggt 3961 tcccgtgtcc accctgagac ttatgagtgg gctaggaaga tggcagtgga tgccctggaa 4021 tacgatgaat cagccgagga tgccaatcct gcaggagccc ttgaagaaat cttggaaaac 4081 ccagagcgac tgaaagacct ggaccttgat gcctttgcag aagagctgga gaggcagggc 4141 tatggtgaca aacacatcac actctatgac atccgggcag agctgagctg tcgatataag 4201 gacctccgga cagcctaccg ctctcccaac acagaggaga tcttcaatat gttaaccaaa 4261 gaaacaccag agaccttcta cattggaaag ctcatcatct gcaatgtcac tggcattgcc 4321 cacaggcgtc cccagggtga gagctatgac caggcgatcc gcaatgatga gacagggctg 4381 tggcagtgcc ccttctgtca gcaggacaat ttccctgaac taagcgaggt gtggaaccac 4441 tttgacagcg gttcgtgccc aggccaggcc atcggtgtca aaacacggct agacaatggt 4501 gtcaccggct tcatccccac caaattcctc agtgacaaag tggtaaagcg gccagaagaa 4561 cgagtgaagg tgggaatgac tgttcactgc cgcatcatga agattgacat tgagaagttc 4621 agtgcagacc tgacctgccg cacctcagac ctcatggaca ggaacaatga gtggaagctg 4681 cccaaagaca cctactatga ctttgatgct gaagctgcag accacaagca ggaggaggac 4741 atgaagcgga agcagcagcg gaccacatac atcaagagag tgatcgcaca cccatccttc 4801 cataatatca atttcaagca agcagaaaag atgatggaga ccatggacca gggtgatgtg 4861 attatccgac caagcagcaa gggcgagaac cacctgacag tgacctggaa agtcagtgat 4921 ggcatctacc agcatgtgga tgtgcgggag gagggcaagg aaaatgcctt cagcctggga 4981 gccactctgt ggatcaacag tgaggaattc gaagatttgg atgagattgt tgctcgctat 5041 gtccagccca tggcatcctt tgcccgggac cttctgaatc acaagtatta tcaggactgc 5101 agcggtgggg accgcaagaa attagaggag ctgctcatca aaactaagaa ggagaagccc 5161 accttcatcc cttatttcat ctgtgcctgc aaggaactgc ccggcaagtt cctactggga 5221 taccagcccc ggggtaaacc caggatagaa tatgtaacgg tgactccaga gggattccgg 5281 taccggggcc agatcttccc aaccgtgaat ggactgttta gatggtttaa ggatcactac 5341 caggatcctg taccaggcat cacccctagc agcagcagca ggacccggac acctgcctct 5401 atcaatgcta ccccagccaa catcaacctt gc-agatctga cacgggctgt gaatgccctg 5461 cctcagaaca tgacttcaca gatgttcagt gccattgctg cggtgacagg ccaaggacag 5521 aaccctaatg ccaccccagc ccagtgggcc tccagccagt acgggtatgg cggcagtgga 5581 ggcggcagca gtgcttacca cgtattccca acgccagccc agcagccagt ggccacacca 5641 ctaatgaccc ctagctactc ctacacgacc ccaagccagc ccatcaccac ccctcagtac 5701 caccagctcc aggccagcac caccccacag tcggcccagg cccagcccca gccctcttcc 5761 agctcccggc aacggcagca gcagccaaag tccaacagcc atgcagccat cgactgggga 5821 aaaatggcgg agcagtggct gcaggaaaag gaggcagaac ggcggaaaca gaagcagcgg 5881 ctgacacctc ggccctcccc cagccccatg atcgaaagca cccccatgtc cattgctggc 5941 gatgccaccc cactcctgga cgagatggat cggtaggggg cctgctcctc ggactctggt 6001 tacctctgag gctgggaaag gcctggctgc ccactgcctc cctccctgcc cctcctttta 6061 tgtccataaa gtggcgtgaa gtgagacgtt ctctttggtg gtcaacccgg atgggtgaca 6121 ggctggatgg ccttgtgaac ttgagctcag tgtatgctag gcaacaattc tcccgctcca 6181 gaccctcacc gaccacctgt cctgggacca ggctgggagg ggagtgtggc agggaggagg 6241 aagaggaagg tgagaatgag tagaacagtt ttgtattcta ctccctacaa gccattttga 6301 acttctgccc tcaccggact ctgggctgtg actggggcac caaactcagc acatgagtct 6361 cccctagctc tcgtggggag agggatgcta tttattcagt ttggggcagg agggagagga 6421 gggaaagtat ttctgaccct gatgccaaca gccgggtggc tgtccaagca ggattgcagg 6481 ggacacaggg aagcactgcc cagcccctgc ctggctgccc tttcccccct gctgctgcca 6541 ccgcttcctg cctgtcattt gaataaacag tgtttctatt gaaaaaaaaa aaaaaaaaaa 6601 aaa