USE OF NOVEL CYTOKINE RECEPTORS AS BIOMARKERS AND THERAPEUTIC TARGETS IN HUMAN CANCER

Abstract

ythropoietin isoforms are described herein, as well as the encoded isoforms, methods of detecting the same, and methods of screening for and treating cancer.

Description

FIELD OF THE INVENTION

[0001]The present invention concerns nucleic acids encoding erythropoietin receptor isoforms, proteins encoded by such nucleic acids, antibodies that bind to such proteins, and methods of using the same.

BACKGROUND OF THE INVENTION

[0002]Erythropoietin (Epo) is the principal hematopoietic growth factor that promotes the viability, differentiation and proliferation of mammalian erythroid progenitor cells (S. Krantz, Blood 77, 419-34 (1991)). The biologic effects of Epo are mediated via its interaction with its specific transmembrane receptor, EpoR (H. Youssoufian, Blood 81, 2223-36 (1993)). The EpoR lacks intrinsic tyrosine kinase activity and upon ligand binding activates a receptor-associated tyrosine kinase Jak2 which is critical for anti-apoptosis and mitogenic signaling via the EpoR (O. Miura et al., Blood 84, 1501-7 (1994); B. Witthuhn et al., Cell 74, 227-36 (1993); J. Ihle, Nature 337, 591-4 (1995); H. Zhuang et al., J. Biol. Chem. 270, 14500-4 (1995)). Activated Jak2 then phosphorylates a number of cytoplasmic proteins as well as the EpoR itself. Expression of Epo receptors has been reported on several non-hematopoietic cell types including vascular endothelial cells, placental tissue, neuronal cells, kidney and cardiomyocytes (A. Anagnostou et al., Proc. Natl. Acad. Sci. USA 91, 3974-8 (1994); S. Masuda et al., J. Biol. Chem. 268, 112-8-16 (1993); S. Sawyer et al., Blood 74, 103-9 (1989); M. Wald et al., J. Ce. Physiol. 167, 461-8 (1996)).

[0003]Recombinant human Epo (r-HuEpo) has been widely used in many different types of cancers for the treatment or prevention of chemo-radiotherapy induced anemia (A. Moliterno and J. Spivak, Hematol. Oncol. Clin. North Am. 10, 345-63 (1996)). For instance, in patients with breast cancer, r-HuEpo has been investigated in clinical trials for its potential beneficial effects in the prevention or treatment of chemotherapy or radiation therapy-related anemia (L. Del Mastro et al., J. Clin. Oncol. 15, 2715-21 (1997); H. Ludwig et al., Ann. Oncol. 4, 161-7 (1993); P. Sweeney et al., Br. J. Cancer 77, 1996-2002 (1998); S. Vijayakumar et al., Int. J. Radiat. Oncol. Biol. Phys 26, 721-9 (1993)), for mobilization of peripheral blood progenitor cells (C. Waller et al., Bone Marrow Transplant 24, 19-24 (1999)), to increase the rate of hematopoietic recovery following high dose chemotherapy (P. Benedetti Panici et al., Br. J. Cancer 75, 1205-12 (1997); S. Filip et al., Neoplasma 46, 166-72 (1999)) as well as use in ex vivo expansion strategies of stem cells (C. Bachier et al., Exp Hematol. 27, 615-23 (1999); L. Pierelli et al., Exp. Hematol. 27, 416-24 (1999); P. Stiff et al., Blood 95, 2169-74 (2000); W. Vogel et al., Blood 86, 1362-7 (1996)). Similarly, r-HuEpo has been investigated in several clinical trials of squamous cell cancers of head-neck (F. Dunphy et al., Cancer 86, 1362-7 (1999); M. Henke et al., radiother Oncol 50, 185-90 (1999); G. Mantovani et al., Oncol. Rep. 6, 421-6 (1999)) and uterine cervix (K. Dusenbery et al., Int. J. Radiat. Oncol. Biol. Phys. 29, 1079-84 (1994)).

[0004]In view of the foregoing, it would be extremely desirable to understand the association of Epo with tumor growth and how EpoR may be involved in cancer pathophysiology and progression.

SUMMARY OF THE INVENTION

[0005]A first aspect of the present invention is an isolated nucleic acid encoding erythropoietin isoform 1, erythropoietin isoform 2, erythropoietin isoform 3, erythropoietin isoform 4, or erythropoietin isoform 5, or a nucleic acid that encodes the opposite or complementary strand of a nucleic acid as set forth above (e.g., a DNA encoding an RNA).

[0006]A second aspect of the present invention is a protein encoded by a nucleic acid as described above (e.g., an isolated and/or purified protein).

[0007]A third aspect of the present invention is an antibody that selectively or specifically binds to a protein as described above.

[0008]A further aspect of the present invention is an oligonucleotide probe that selectively or specifically binds to a nucleic acid as described above.

[0009]A further aspect of the present invention is a method of screening a subject for cancer, comprising: detecting the presence or absence of a nucleic acid encoding an isoform as described above in the subject, the presence of such a nucleic acid indicating the subject is afflicted with or at risk of developing cancer.

[0010]A further aspect of the present invention is a method of screening a subject for cancer, comprising detecting the presence or absence of a protein or isoform as described above in the subject, the presence of such a protein indicating the subject is afflicted with or at risk of developing cancer.

[0011]Particular cancers which may be screened by the methods described herein include, but are not limited to, breast, cervix, ovarian, prostate, colon and lung cancer.

[0012]The foregoing and other objects and aspects of the present invention are explained in detail in the drawings herein and the specification set forth below.

BRIEF DESCRIPTION OF THE DRAWINGS

[0013]FIG. 1. The organization of the EpoR gene (GenBank accession number S45332, SEQ ID NO: 1). The splicing that results in the mature mRNA for the wild-type receptor (SEQ ID NO: 3), and five alternatively spliced isoforms (1-5) described herein are depicted schematically. The translated regions of the gene are indicated in black, whereas untranslated regions are indicated in white. Novel amino acid translations that result from alternative splicing of the EpoR gene transcript are indicated in grey.

[0014]FIG. 2. Changes in the open reading frames (ORFs) of mature mRNA sequences from the full-length wild-type receptor in the isoforms of EpoR described herein. Isoform 1 (SEQ ID NO: 4): Additional nucleotides from intron 6 (nucleotides 5949-6062. SEQ ID NO: 1) are spliced between exons 6 and 7. Isoform 2 (SEQ ID NO: 6): Splicing at the 5' end of exon 8 occurs 19 nucleotides upstream (nucleotide 7498) from that seen in the full-length wild-type message (nucleotide 7517). Isoform 3 (SEQ ID NO: 8): Intron 7 is not spliced out of the final message. Isoform 4 (SEQ ID NO: 10): Intron 5 is not spliced out of the final message. Isoform 5 (SEQ ID NO: 12): exon 6 is skipped, with exon 5 spliced directly to exon 7. Putative C-terminal amino acid sequence changes from wild-type EpoR are depicted in bold.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015]The present invention is explained in greater detail below. This description is not intended to be a detailed catalog of all the different ways in which the invention may be implemented, or all the features that may be added to the instant invention. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure which do not depart from the instant invention. Hence, the following specification is intended to illustrate some particular embodiments of the invention, and not to exhaustively specify all permutations, combinations and variations thereof.

[0016]Nucleic acid as used herein refers to any type of nucleic acid, including naturally occurring and synthetic nucleic acids and including both DNA and RNA.

[0017]Subjects with which the present invention may be carried out are generally mammalian subjects, including both human subjects and non-human subjects (e.g., dog, cat, horse, rabbit, rat) for veterinary or research purposes.

[0018]Any type of antibody may be used in the present invention. The term "antibodies" as used herein refers to all types of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE. Of these, IgM and IgG are particularly preferred. The antibodies may be monoclonal or polyclonal (with monoclonal antibodies preferred) and may be of any species of origin, including (for example) mouse, rat, rabbit, horse, or human. See, e.g., M. Walker et al., Molec. Immunol. 26, 403-11 (1989). Antibody fragments that retain specific binding to the protein or epitope bound by the antibody are included within the scope of the term "antibody" and include, for example, Fab, F(ab')₂, and Fc fragments, and the corresponding fragments obtained from antibodies other than IgG. Such fragments can be produced by known techniques. The antibodies may be chimeric or humanized, particularly when they are used for therapeutic purposes.

[0019]Applicants specifically intend that all United States patent references cited herein be incorporated herein by reference in their entirety.

1. Nucleic Acids.

[0020]As noted above, a first aspect of the present invention is a nucleic acid encoding an erythropoieitin receptor isoform as described herein. In certain embodiments the nucleic acid may be an RNA such as an mRNA, or may be a DNA.

[0021]In one embodiment, the nucleic acid encodes erythropoietin receptor isoform 1 and has the sequence given herein as SEQ ID NO: 4.

[0022]In another embodiment, the nucleic acid encodes erythropoietin receptor isoform 2 and has the sequence given herein as SEQ ID NO: 6.

[0023]In another embodiment, the nucleic acid encodes erythropoietin receptor isoform 3 and has the sequence given herein as SEQ ID NO: 8.

[0024]In another embodiment, the nucleic acid encodes erythropoietin receptor isoform 4 and has the sequence given herein as SEQ ID NO: 10.

[0025]In another embodiment, the nucleic acid encodes erythropoietin receptor isoform 5 and has the sequence given herein as SEQ ID NO: 12.

[0026]In another embodiment, the nucleic acid that encodes the opposite strand of a nucleic acid as set forth above (e.g., is a DNA encoding an RNA).

[0027]Nucleic acids as described above may be natural or synthetic, and can be produced in accordance with techniques known in the art or variations thereof which will be apparent in light of the disclosure herein.

[0028]Nucleic acids as described above may be coupled to appropriate regulatory elements such as a promoter to produce a recombinant nucleic acid construct, which construct may be inserted into a host cell in which the promoter is operable so that the encoded protein is expressed by the host cell. Recombinant techniques and the production of proteins in recombinant cells may be carried out in accordance with known techniques.

2. Antibodies.

[0029]Polyclonal antibodies used to carry out the present invention may be produced by immunizing a suitable animal (e.g., rabbit, goat, etc.) with the antigen to which the monoclonal antibody binds, collecting immune serum from the animal, and separating the polyclonal antibodies from the immune serum, in accordance with known procedures. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Corynebacterium parvum are especially preferable.

[0030]Monoclonal antibodies of the present invention may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique (Kohler, G. et al. (1975) Nature 256:495-497; Kozbor, D. et al. (1985) J. Immunol. Methods 81:31-42; Cote, R. J. et al. (1983) Proc. Natl. Acad. Sci. 80:2026-2030; Cole, S. P. et al. (1984) Mol. Cell. Biol. 62:109-120). Briefly, the procedure is as follows: an animal is immunized with antigen or immunogenic fragments or conjugates thereof. For example, haptenic oligopeptides of antigen can be conjugated to a carrier protein to be used as an immunogen. Lymphoid cells (e.g. splenic lymphocytes) are then obtained from the immunized animal and fused with immortalizing cells (e.g. myeloma or heteromyeloma) to produce hybrid cells. The hybrid cells are screened to identify those which produce the desired antibody.

[0031]Human hybridomas which secrete human antibody can be produced by the Kohler and Milstein technique. Although human antibodies are especially preferred for treatment of human, in general, the generation of stable human-human hybridomas for long-term production of human monoclonal antibody can be difficult. Hybridoma production in rodents, especially mouse, is a very well established procedure and thus, stable murine hybridomas provide an unlimited source of antibody of select characteristics. As an alternative to human antibodies, the mouse antibodies can be converted to chimeric murine/human antibodies by genetic engineering techniques. See V. T. Oi et al., Bio Techniques 4(4):214-221 (1986); L. K. Sun et al., Hybridoma 5 (1986).

[0032]In addition, techniques developed for the production of "chimeric antibodies", the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity can be used (S. L. Morrison, et al. Proc. Natl. Acad. Sci. 81, 6851-6855 (1984); M. S, Neuberger et al., Nature 312:604-608 (1984); S. Takeda, S. et al., Nature 314:452-454 (1985)). Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce isoform-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobin libraries (D. R. Burton, Proc. Natl. Acad. Sci. 88, 11120-3 (1991)).

[0033]Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature (R. Orlandi et al., Proc. Natl. Acad. Sci. 86, 3833-3837 (1989)); G. Winter et al., Nature 349, 293-299 (1991)).

[0034]Antibodies that selectively bind to a particular erythropoietin receptor isoform as described herein (i.e., that selectively bind to one of isoforms 1-5 but do not bind to the other of isoforms 1-5) can be identified in accordance with known techniques, such as their ability to compete with labeled antibody to in binding to that isoform in a competitive binding assay.

[0035]If desired, antibodies specific for a particular isoform can be used to produce anti-idiotypic (paratope-specific) antibodies. See e.g., McNamara et al., Science 220, 1325-26 (1984), R. C. Kennedy, et al., Science 232, 220 (1986).

3. Immunoassay Techniques.

[0036]Those skilled in the art will be familiar with numerous specific immunoassay formats and variations thereof which may be useful for carrying out the method disclosed herein. See generally E. Maggio, Enzyme-Immunoassay, (1980)(CRC Press, Inc., Boca Raton, Fla.); see also U.S. Pat. No. 4,727,022 to Skold et al. titled "Methods for Modulating Ligand-Receptor Interactions and their Application," U.S. Pat. No. 4,659,678 to Forrest et al. titled "Immunoassay of Antigens," U.S. No. 4,376,110 to David et al., titled "Immunometric Assays Using Monoclonal Antibodies," U.S. Pat. No. 4,275,149 to Litman et al., titled "Macromolecular Environment Control in Specific Receptor Assays," U.S. Pat. No. 4,233,402 to Maggio et al., titled "Reagents and Method Employing Channeling," and U.S. Pat. No. 4,230,767 to Boguslaski et al., titled "Heterogenous Specific Binding Assay Employing a Coenzyme as Label."

[0037]Antibodies as described herein may be coupled or conjugated to a solid support suitable for a diagnostic assay (e.g., beads, plates, slides or wells formed from materials such as latex or polystyrene) in accordance with known techniques, such as precipitation. Antibodies as described herein may likewise be coupled or conjugated to detectable groups such as radiolabels (e.g., ³⁵S, ¹²⁵I, ¹³¹I) enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase), fluorescent labels (e.g., fluorescein), chemiluminescent labels (e.g., acridinium groups, metalloporphyrins such as phthalocyanine dyes, luminol, etc.), metal atoms (e.g., technetium-99m), etc., in accordance with known techniques. See, e.g., U.S. Pat. No. 4,472,509 to Gansow (metal chelates to monoclonal antibodies); U.S. Pat. No. 5,061,641 to Schochat et al.; and U.S. Pat. No. 4,861,869 to Nicoleotti et al. (radiolabelling proteins).

[0038]Immunoassays, or other types of assays to detect and/or quantitate the level of the isoform in samples as described below, may be used in screening assays to detect pathologic states associated with aberrant levels of isoform expression (e.g., tumors, inflammatory states), diagnostic studies, prognostic studies, or to monitor the progression or diminution of isoform expression in correlation with disease state.

[0039]Samples that may be collected for use in carrying out the immunoassay may be tissue samples from the organ or tissue of interest within the subject, such tissue generally of most interest being those types of tissues/cells that express differing amounts of isoform in pathologic states as compared to non-pathologic states, or biological fluids such as blood (including blood fractions such as blood plasma or blood serum), urine, cerebrospinal fluid, etc). Examples may include overexpression or aberrant expression of the isoform in various types of malignancies (e.g ovarian cancer, endometrial cancer, pancreatic cancer, breast cancer, urinary bladder cancer, lung cancer, etc.), as well as overexpression or aberrant expression in other pathologic states.

[0040]A biological sample may be a cell sample, with an intervening culturing step being performed between the time the cell sample is collected from the subject and the immunoassay is carried out on the biological sample.

[0041]For immunohistological techniques, a tissue sample is collected from the subject, and the presence or absence of binding of an antibody of the invention is detected. The presence of binding of the antibody in an abnormal pattern or a pattern indicative of a tumor or cancer indicates the presence of a tumor or cancer in the subject from which the tissue sample is collected. The presence of the antigen in a metastatic tumor deposit can also be used to determine a likely source of the primary tumor. Any suitable immunohistology format may be used. The tissue sample may include patient biopsies, resections or cells for cytologic study. A similar technique to immumohistogy is the use of similar techniques to detect and/or phenotype cells in body fluids or other suspensions as is used for flow cytometric examination.

[0042]For in vivo diagnostic purposes the antibody according to the invention is coupled to or provided with a suitable externally detectable label, such as e.g. a radiolabel as described above or a metal atom (e.g., technetium-99m), and administered to a subject (e.g., by intraveneous or intraarterial injection), in an amount sufficient to produce an externally detectable signal, whereupon the possible localized accumulation of antibody in the body is determined, with a localized accumulation of the antibody (in a region other than that which would ordinarily be expected for normal subjects or subjects free of disease) indicating the present of a tumor in that subject.

4. Nucleic Acid Assay Techniques.

[0043]Detection of mRNAs specific to EpoR isoforms 1, 2, 3, 4, and 5 may be carried out by any suitable technique, including but not limited to using reverse transcriptase-polymerase chain reaction (RT-PCR) amplification with isoform-specific primers and Southern blot analysis of the resulting RT-PCR amplicons. For example, PolyA.sup.+ RNA may be isolated by any technique known by those skilled in the art from patients patient cells and/or cancer cells, including but not limited to breast, colon, lung, ovary, and prostate cells or cancer cells. Methods for RT-PCR amplification of the isolated RNA are known in the art and may be carried out using EpoR isoform-specific primer pairs, preferably as described below.

[0044]Oligonucleotide probes (or primers) that specifically bind to a nucleic acid encoding an isoform as described above (including the opposite strands thereof), and pairs of probes (where at least one member of the pair is specific for a nucleic acid encoding one particular isoform), are also an aspect of the present invention. In general, such probes are from 8 or 10 nucleic acids in length up to 40 or 50 nucleic acids in length, or more. By "specifically bind" is meant that a probe binds to a nucleic acid (or complement thereof) that encodes one isoform as described herein, but does not bind to a nucleic acid (or complement thereof) that encodes another isoform as described herein. Probes may optionally be labeled with a detectable group such as a radioisotope, enzyme, or member of a binding pair in some assay formats. Where a pair of probes or primers is used for amplification, it will be appreciated that only one member of the pair need be isoform-specific, and that the other member of the pair may be one which will bind to nucleic acids encoding more than one of the isoforms described herein, so long as the primer pair specifically amplifies only nucleic acid encoding one of the isoforms described herein. Examples of such oligonucleotide probes, and pairs thereof, are as follows:

TABLE-US-00001 Primer pair specific for intron 6 insert (isoform 1) (SEQ ID NO: 14) 28AS-: 5' TCA AGC GGC TGG TTC CTT CGA A 3' (SEQ ID NO: 15) ER4-5: 5' GCA GGG AGC GTA CAG AGG GTG GAG 3' Primer pair specific for intron 7 insert (isoform 2) (SEQ ID NO: 16) 33AS: 5' GAA GAA ATA GCA CCA ACC TGG AAG 3' (SEQ ID NO: 17) 31S: 5' CTG ACG CCT AGC GAC CTG GAC C 3' Primer pair specific for intron 7 unspliced (isoform 3) (SEQ ID NO: 18) 31AS: 5' GCA GTT TGG CTG CAA GAA GCA 3' (SEQ ID NO: 17) 31S: 5' CTG ACG CCT AGC GAC CTG GAC C 3' Primer pair specific for intron 5 unspliced (isoform 4) (SEQ ID NO: 19) 26S: 5' GGA GCC AGG GCG AAT CAC GG 3' (SEQ ID NO: 20) 32S: 5' GCC TTC AAA CTC GCT CTC TG 3' Primer pair specific for exon 6 skipped (isoform 5) (SEQ ID NO: 21) 34AS 5' GCT TCA GAG CCC GCT AGG CGT 3' (SEQ ID NO: 15) ER4-5 5' GCA GGG AGC GTA CAG AGG GTG GAG 3'

Note that, in the foregoing pairs, the primers of SEQ ID NO. 14, 16, 18, 19 and 21 are specific for the identified isoform, and the primers of SEQ ID NO. 15, 17 and 20 are not specific. In each pair, only one primer need be specific to provide an isoform-specific primer pair.

[0045]Blotting techniques are well known in the art. See, e.g., Sambrook et al., Molecular Clooning: a Laboratory Manual 3rd Ed. (Cold Spring Harbor, N.Y.); Ausubel et al. Current Protocols in Molecular Biology (Green Publishing Associates, Inc. and John Wiley & Sons, Inc., New York). The nucleic acids resulting from RT-PCR amplification may be separated by gel electrophoresis and immobilized on a suitable matrix, e.g. a filter of nitrocellulose. The presence of target sequences among the amplification products may be shown by incubation of the blotted amplicons with a probe (usually labeled) under conditions that promote denaturation and rehybridization. Because the probe is designed to base pair with target sequences, the probe will bind under renaturing conditions. Unbound probe is then removed, and detection of target sequences may be accomplished via known techniques to detect the labeled probe.

[0046]The present invention and the various methods and compounds therein are explained in greater detail in the following non-limiting examples.

Example 1

[0047]The Erythropoietin Receptor (EpoR) gene. The human EpoR gene has been cloned and sequenced as previously described (Noguchi et al. (1991) Blood 78:2548-2556). The gene spans 8.6 kilobases, and comprises of 8 exons with 7 intervening introns, the latter of which range in size from 81 by to 2.1 kb. The organization of the EpoR gene is outlined in FIG. 1. The full-length wild-type form of EpoR comprises of 508 amino acids (SEQ ID NO: 2) in three domains: extracellular, transmembrane (TM), and cytoplasmic. Exons 1-5 encode for the extracellular domain of EpoR, exon VI encodes for the transmembrane domain, while exons VII and VIII encode for the cytoplasmic domain of the receptor. In the examination of EpoR expression in tumor vasculature, analysis by RT-PCR indicated a high level of EpoR mRNA expression in breast cancer cells, as well as squamous cell cancers of head-neck and uterine cervix was observed.

Example 2

[0048]Novel Isoforms of EpoR mRNA Transcripts. The resulting RT-PCR amplification products derived from human cervix, breast, prostate, and ovarian cancer cell lines were sequenced and analyzed. The results of this study revealed five alternatively spliced EpoR mRNA transcripts that differ from the mature, full-length wild-type EpoR mRNA. Using isoform-specific PCR primers, transcripts corresponding to each isoform were detected in breast, colon, lung, ovarian and prostate cancer. The organization of these five isoforms is outlined in FIG. 1. The alternative forms of EpoR predicted to be coded for from these alternatively spliced mRNAs fall into two categories. Isoforms 1, 2, and 3 are described as truncated (EpoR-T), and possess the extracellular and transmembrane domains of the wild-type receptor, while lacking portions of the cytoplasmic domain. Isoforms 4 and 5 are described as soluble (EpoR-S), and only possess the extracellular domain of the wild-type receptor intact. The changes in the putative C-terminal amino acid sequence encoded by these mRNAs are outlined in FIG. 2.

Example 3

[0049]EpoR Isoform 1. The mRNA that codes for Isoform 1 contains an additional 114 nucleotides from intron 6 (nucleotides 5949-6062, SEQ ID NO: 1) spliced between exons 6 and 7. The resulting mRNA will code for an EpoR peptide 285 amino acids in length (SEQ ID NO: 5) with a severe truncation in the cytoplasmic region. At the C-terminal, 9 novel amino acids (M V R E G S R R R STOP) inserted at position 277 of the full-length EpoR peptide sequence.

Example 4

[0050]EpoR Isoform 2. The mRNA that codes for Isoform 2 is the result of an alternative splicing event between the 3' end of exon 7 and 5' end of exon 8, in which an additional 19 nucleotides (nucleotides 7498-7516, SEQ ID NO: 1) are added to the 5' end of exon 8. The mRNA from this splicing event codes for an EpoR peptide 317 amino acids in length (SEQ I NO: 7) with a severe truncation in the cytoplasmic domain, in which 12 novel amino acids (V G A I S S A V A V P E STOP) are inserted at position 306 of the EpoR peptide sequence. As with Isoform 1, Isoform 2 also possesses a truncation of the cytoplasmic domain of the full-length peptide sequence of EpoR.

Example 5

[0051]EpoR Isoform 3. The translation of isoform 3 results from a processed EpoR mRNA in which sequences from intron 7 (nucleotides 7422-7516, SEQ ID NO: 1) are not spliced out of the final message. The resulting translation is a 328 amino acid peptide (SEQ ID NO: 9), with 23 novel amino acids introduced to the C-terminus (V G G L V V P S V P G L P C F L Q P N C R P L STOP) at position 306 of the EpoR peptide sequence. As with Isoforms 1 and 2, Isoform 3 possesses a truncation of the cytoplasmic domain of the full-length peptide sequence of EpoR. The sequence of the ORE of the mRNA message (SEQ ID NO: 8) and the peptide sequence of EpoR Isoform 3 (SEQ ID NO: 9) is identical to the translation predicted from an mRNA described previously (Nakamura et al. (1992) Science 257:1138-1141).

Example 6

[0052]EpoR Isoform 4. The processed EpoR mRNA that translates into Isoform 4 contains sequences from intron 5 (nucleotides 5061-5144, SEQ ID NO: 1) are not spliced out of the final message. The resulting translation is a 267 amino acid peptide (SEQ ID NO: 11) with 21 novel amino acids (G E A P G G G V G G A R A N H G A S P P P STOP) introduced to the C-terminus at position 247 of the full-length EpoR peptide sequence. This isoform of EpoR possesses neither the transmembrane nor cytoplasmic domains of the full-length receptor. The translation that codes for Isoform 4 results in a soluble form of EpoR, containing the extracellular domain of the receptor only.

Example 7

[0053]EpoR Isoform 5. Isoform 6 is a translation that results from the alternatively processed EpoR mRNA in which sequences from exon 6 are skipped, i.e. exons 5 and 7 are spliced together directly. The translation of this message results in a 248 amino acid peptide (SEQ ID NO: 13), in which 2 novel amino acids (G L STOP) are introduced at position 247 of the full-length peptide sequence of EpoR. As with Isoform 4, Isoform 5 of EpoR is a soluble form of the receptor that comprises of only the extracellular domain.

[0054]The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Sequence CWU 1

2118647DNAHomo sapiensCDS(1916)..(2030)exon 1 1ggatccaccc acctcggcct cccaaagtgc tgggattaca ggcatgagca ctgtgcatgg 60actatttatt tatttttttg aaacagagtt tcaatcttgt tgcacagcct ggagtgcaat 120ggtgtgatct cagctcactg caacctctgc cttctggttt caagcaattc tcctgcctca 180gcctcctgag tagctgggat tacaggcacc caccaccacg ctcgaatata tatatatatt 240ttttgagacg gagtccgctc tgtcaccagg ctggagtgca gtggccaaat atcggctcac 300tgaaacctcc ggctcctggg ttcaagcgat tctcctgcag cctcccaagt agctgggatt 360acaggcatgc agcaccacgc ccatctaatt tttgtatttt tggtagagat ggggttttac 420catgttggcc aggatggtct tgatctcttg acctcgtgat ctgcccacct cggcctccca 480aagtgctggg attacaggcg tgacgaccgc gcccggccta cgcctggcta atttttgtat 540ttttagtaga gacgtggttt cgccatgttg cccaggctgg tctcgaactc ctgacctcat 600gatccgcctg tctcggcctc ccaaagtgtt gggattacaa gtatgagcca ccgcgccact 660agccaatttt ttttattttt tgagatgcag tctcactctg ttgcccaggc tggagttgca 720gtggcatgat cttggctcac tgcaatcttc atctcccaga ctgaagcagt tctcatgcct 780cagcctcctg agtagctggg attacagcac acgccaccac acctggctaa tttttgtatt 840tttagtagag atgggatttc accatgttgg ccaggctggt ctcaaactcc tgacctcaag 900tgatttgccc acgtcggcct cccaaagtgc tgggattata ggcgtgagcc accgcccagc 960ccaagagaat aaaaatgtgg gtggtaaaaa tttttttccc aaaaattcgt aaatgaaaat 1020ctcacatatt atgcatactg cccaggagca tggcctagca ctgtgcaaac actcaactgc 1080tggtcgttgc aaggattatt attggccggc ttcagtggct tgctggtatt cccagcacat 1140tgggagatgg aggctggagg attgcttaag tccgggattt caagaccagc ctggacaaca 1200tagtgggatc ccatctctac aaagaatttt aaaaattagc caggtgcagt gggaagattg 1260cttcagtcca gaggctgcag tgagctatga ttgtgccact gcactccagc ctgggtgaca 1320gagcaacacc ctgagacaga gagagagagg gggaaggagg gaaggaggga aggaaggaag 1380gaaggaagga aggaaggaag gaaggaagga aggaaggaaa ggagagagag agagagagag 1440agagagagag agagagaaaa taatttttat ttatttccag gctgggaaga gatgctgatt 1500tctgcgataa aatcagtagg tacatttttt ggaatgttcg ctatgtgcca ggctagattt 1560tacagatgag aagtctgaag ctcaggtaag gtaagtcacc tgtccagggc cacaaagaaa 1620aaaaaaacgt gtgtctgaag ccagaacggg agctgttgcg cccaactccc tcccctgccc 1680ccaagcggcc tctgggctcg ggaagggccc ctgcctcctc ccgccaggca cttatctcta 1740cccaggctga gtgctggccc cgcccctcgg ggatctgcca cttagaggcg cctggtcggg 1800aagggcctgg tcagctgcgt ccggcggagg cagctgctga cccagctgtg gactgtgccg 1860ggggtggggg acggaggggc aggagccctg ggctccccgt ggcgggggct gtatc atg 1918 Met 1gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt ctc 1966Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys Leu 5 10 15ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac ccc 2014Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp Pro 20 25 30aag ttc gag agc aaa g gtaaggatga gctgcgtgtg gacccctacg ctggagcctg 2070Lys Phe Glu Ser Lys 35caggaccatg ctggggcctg aactcccagc ctaggtcctg ggggccatgc tgtttctgga 2130cttcctgacc gggtcctggg ggccaagctg gcatctgaac ccttagactg ggtcctggat 2190gggtgggggg cggggtgggg tatgttagga tccaagactc ctgatcgcgt cccgggcaag 2250agctagagtg ggcttaacat tcccgtttta ccttttcagg gagtctggga catgctaaat 2310cctaaggggg ctgacttggt gctaaggtcc ctggggggtg gggaccaagc cgatccctag 2370gggagggagg gtaaagcccg ggtccgagtt agagggccaa gccacaggct actgtaaaca 2430cggtttgtgt gagggcgcca gatcacttgc ccggcccggt ggagggaggg aggcgggggg 2490cacggttggc gctatcggtt ggcggggagc ctgccggggc cgataggggg cccgcctctc 2550cgcacacacc cccagccgcg cgcgtgtcct aggctggggc ggggctggca gtcccgagct 2610cgaggtcttg aacgccgcgc ccagctcagc tggccgctgg gtgggcaggt gtgcgccagt 2670ggtgcacggc gggggacagt aaggcgagaa acttgcccct gggaattagg ggggcaccac 2730ctctgcggac ccctccaagg gacccgcttg ggaagatggc agggcggggc ttttttctta 2790tcgggtccgc ccaggctgcg ggagggaaga ggagggggct gtctcccgag gatagagctc 2850agacccccat gcccttcctt tgtcgcccct ccccag cg gcc ttg ctg gcg gcc 2903 Ala Ala Leu Leu Ala Ala 40cgg ggg ccc gaa gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg 2951Arg Gly Pro Glu Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu45 50 55 60gtg tgt ttc tgg gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac 2999Val Cys Phe Trp Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn 65 70 75tac agc ttc tcc tac cag ctc ga gtgagtccga tccggcgggt gcctccaagg 3052Tyr Ser Phe Ser Tyr Gln Leu Glu 80gcggagggag ggggtggggc agagctccct ggaggtcgta gcctcgtatg tcccctgctg 3112tttgaggccc gacggcgcct ccagtcgtgg tcactggagg gaaacctgcg ggtccagggc 3172tggcacgcct ctatgggccg gggcgcgaac actcccgcga tcaccgctgg aacgcgaccc 3232caaacatcag gctgggataa caacgcctcc aaatcgaggg taaggcgtta ctacgtcggg 3292gctgggacgc cttctcgagg tagtatccaa aaggaggcca gcagtgctca tgcctgtaat 3352cccaactctt tggaaggtcg agcggaagaa ccgcttgagc ccaggtgttc aagaccagcc 3412tgggcaacac agcgagatcc ccgtctctta aaaaaaaatt agactgggcg cggctgcacg 3472cctgtaatcc cagcactttg ggaggctgag gcgggcggat cacctgaggt cgggagtttg 3532agagccagcc tggccaacat ggagaaactc tatctctact aaaaatacaa aattagccgg 3592gcgtggtggc gcatgcctgt gatcccagct actcgggagg ctgaggcagg agaatcgctt 3652gaacccggga ggcggaggtt gcggtgagcc gaggtagcgc cattgcactc cagcctgggc 3712aacaagagcg aaactccgtc tcaaaaaaaa aaaaaataaa agccaggcgt ggcgcgtgcc 3772tgtggtctca actacttggg aagctgaggt gggaggatcc cttaagcccc agaatttgag 3832gctgcagtga gccatgatcg cgccactgca ctccagcctg ggcgacgaag gaacaccttg 3892tcacacacac acacaaggct agaccttgtg tcacacatac acactgcccc ccacaggccg 3952ggcaatgcca actccccggt cccccctccc aacctgctcc cttccctggg cgcatag g 4010gat gag cca tgg aag ctg tgt cgc ctg cac cag gct ccc acg gct cgt 4058Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala Pro Thr Ala Arg85 90 95 100ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc gac acg tcg agc 4106Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala Asp Thr Ser Ser 105 110 115ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc ggc gct ccg cga 4154Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser Gly Ala Pro Arg 120 125 130tat cac cgt gtc atc cac atc aat gaa gta g gtaagtgctc tgggaatgga 4205Tyr His Arg Val Ile His Ile Asn Glu Val 135 140ggagtggtcg gaggagaggg tctcagtcct cgcccacctg accaaccccc atgcctgcag 4265tg ctc cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag 4312Val Leu Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu 145 150 155agc ggc cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg 4360Ser Gly His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met 160 165 170acg tct cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca 4408Thr Ser His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala175 180 185 190ggg agc gta cag agg gtgaggccag cccctacggc ccagccccca aagctccact 4463Gly Ser Val Gln Arg 195gactacggcc cagccacgcc tctcgaggtc gcgcccggtg ccgctttcag ggccggtccg 4523taacatccca catcccatta ccctggtgct gaagaccgtt ccacgcccac agacacagcc 4583ccctttccta atgtcctcgc aagcctgttg aaccccaact tcttctccct ccggcccgta 4643accctagacc cctttagcgc ccgggtccct ctacgagtgc tagcccagat attaaattgc 4703ccgggtcccg ccctttcgta ccagagactc tctctctgat tggccctgag ctttcttggg 4763ctcctccccc tactcttatt ggtcccattg caattctagg gcaccgtttt cctttcccct 4823gattggctca gttccaccag ggcccgcccc cacgtcatct atttttgtct gctacgcgtc 4883cctcgccctg attccgcccc cag gtg gag atc ctg gag ggc cgc acc gag tgt 4936 Val Glu Ile Leu Glu Gly Arg Thr Glu Cys 200 205gtg ctg agc aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc 4984Val Leu Ser Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg 210 215 220gcg cgt atg gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg 5032Ala Arg Met Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser 225 230 235gag cct gtg tcg ctg ctg acg cct agc g gtgaggcccc aggcgggggt 5080Glu Pro Val Ser Leu Leu Thr Pro Ser 240 245gtaggaggag ccagggcgaa tcacggggca agcccaccgc cctgacctcc tccccgcctc 5140ttag ac ctg gac ccc ctc atc ctg acg ctc tcc ctc atc ctc gtg gtc 5188 Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser Leu Ile Leu Val Val 250 255 260atc ctg gtg ctg ctg acc gtg ctc gcg ctg ctc tcc cac cgc cg 5232Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu Ser His Arg Arg 265 270 275gtgagctccc catttgggcg ctgggcccag actcctcccc gccaacggtc ctctttcact 5292atggaaacct aggctcagag agagacacgc acttgcccaa ggtcacgcag taaggattca 5352catcagtggc agggctggga tgcatgccag actagaccca gactcttcgt taacattttc 5412tgctcttggg gactttcacc tgattttcct tctacatcag gggctgccat ttcttgggtc 5472cctttgttag ttcctttccc cagtgtcatc acctttgtaa aatcaactag atggatttag 5532tgaaagaatt taagaccctg aatgcctccg cacccctgcg gtcaagcttc tcagacacta 5592tgatcagact agccgttctg aggtatttgt aattccaagc acacactagg tggtttcaca 5652cccccaagct tttgcccatg ctgttccctc tgcctggaat gcccttcctg ccttgtctgc 5712taagcaatct tctagtcgtc tttcatggcc ctgttcattt acttggttgg aaaatacaaa 5772cagagtgcca aacatgtgcc aggcactgga gagagaatgg agaacaagct agaccctgac 5832cacaagtccc tgaccttgtg gatctcaagt caacaaacaa gggacccaag aaatatttga 5892tgacaaattg taatgagtga tatcacagaa acaaacagaa tgtggtgaca tgacaggatg 5952gtcagggaag gctccaggag gaggtgacat cagagtggaa acctgaagat tggaaggaag 6012cagccgcttg aaaagtgggg agaagaaaca gcaagtgcaa aggccctgag gtgggaatga 6072gattggaacg ttcagccagc ttcaagaatt gccacatgca tggcctggca tggtggctca 6132cgcctgtaat cccagcactt tgggatgccg aggcaggcag atcacctgag gttgggagtt 6192cgcgaccagc ctgaccaaca tggagaaacc ccacctctac taaaaataca aaactagcca 6252agcgtggtgg cacatgcctg taatccccgc tactcgggag gctgaggcag gagaatcact 6312tgaacctggg aggtggaggt tgcgggtgag ccgagatcgt gccatcgcat tccagcctgg 6372gcaataagag tgaaactccg tctcaaaaaa aaaaaaaaaa ttgccacatg gctagagtgg 6432tatgtaaggg ggtgtggcag atattgagat gagggaggtg acaggggtca tataacgcag 6492ggccttctgc agggtggtgg ggaggagttt ggaatttttt ttttttttga gacagagtca 6552ctcttgtcgc ccaagctgta gtgcagtgca gcagtcttgg ctcactgcaa ctctgcctcc 6612caggttcaag tgattctcct gcctcaaccg cctgagtagc tgagattaca ggcgtgcatg 6672cccggctaat tttgtagttt tagtagagac ggggttccac catgttggcc aggctggtct 6732caaactcctg acctcaggtg atctgctcac atcagcctct caaagtgctg ggattatagg 6792catgagccac cgtgcctggc ttggatttta tcctaaatgc ctctctcatt accccagaag 6852gtaacataat atttatctat gaagtgacat catggacctc ctggaaaaat ctgggccagg 6912gttttgggtt ttttaattta ttttatttta ttttttttag agatgggggt ctcactatgt 6972ttcctaggct ggtcttgaac tcctgggttc aaatgatcct cccacctcag cctcccaaag 7032tactgggatt atagtgctgg tgtaaaccac tgcacctggc catggccagg attaaaggga 7092gaatgaccaa ggtatattga actcctatgc acccttcaat accctgttcc atttaccctt 7152ttgtagggcc ttgctgatgc ttcagccaaa acccctgtcc cctggccctg atgtactcct 7212ctgcctccat tgtgatcaca gggaccaagt gtatctgtgc ctctatgact gggagtggag 7272ggggaattgg tgagtattca atgagtcata tctatgtaac tatttatatt ggcttcaaca 7332g g gct ctg aag cag aag atc tgg cct ggc atc ccg agc cca gag agc 7379 Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser Pro Glu Ser 280 285 290gag ttt gaa ggc ctc ttc acc acc cac aag ggt aac ttc cag 7421Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe Gln 295 300 305gtaggtggcc tggttgtccc ctcagtgcct gggcttccct gcttcttgca gccaaactgc 7481aggcctctct gagcaggttg gtgctatttc ttcag ctg tgg ctg tac cag aat 7534 Leu Trp Leu Tyr Gln Asn 310gat ggc tgc ctg tgg tgg agc ccc tgc acc ccc ttc acg gag gac cca 7582Asp Gly Cys Leu Trp Trp Ser Pro Cys Thr Pro Phe Thr Glu Asp Pro 315 320 325cct gct tcc ctg gaa gtc ctc tca gag cgc tgc tgg ggg acg atg cag 7630Pro Ala Ser Leu Glu Val Leu Ser Glu Arg Cys Trp Gly Thr Met Gln 330 335 340gca gtg gag ccg ggg aca gat gat gag ggc ccc ctg ctg gag cca gtg 7678Ala Val Glu Pro Gly Thr Asp Asp Glu Gly Pro Leu Leu Glu Pro Val 345 350 355ggc agt gag cat gcc cag gat acc tat ctg gtg ctg gac aaa tgg ttg 7726Gly Ser Glu His Ala Gln Asp Thr Tyr Leu Val Leu Asp Lys Trp Leu360 365 370 375ctg ccc cgg aac ccg ccc agt gag gac ctc cca ggg cct ggt ggc agt 7774Leu Pro Arg Asn Pro Pro Ser Glu Asp Leu Pro Gly Pro Gly Gly Ser 380 385 390gtg gac ata gtg gcc atg gat gaa ggc tca gaa gca tcc tcc tgc tca 7822Val Asp Ile Val Ala Met Asp Glu Gly Ser Glu Ala Ser Ser Cys Ser 395 400 405tct gct ttg gcc tcg aag ccc agc cca gag gga gcc tct gct gcc agc 7870Ser Ala Leu Ala Ser Lys Pro Ser Pro Glu Gly Ala Ser Ala Ala Ser 410 415 420ttt gag tac act atc ctg gac ccc agc tcc cag ctc ttg cgt cca tgg 7918Phe Glu Tyr Thr Ile Leu Asp Pro Ser Ser Gln Leu Leu Arg Pro Trp 425 430 435aca ctg tgc cct gag ctg ccc cct acc cca ccc cac cta aag tac ctg 7966Thr Leu Cys Pro Glu Leu Pro Pro Thr Pro Pro His Leu Lys Tyr Leu440 445 450 455tac ctt gtg gta tct gac tct ggc atc tca act gac tac agc tca ggg 8014Tyr Leu Val Val Ser Asp Ser Gly Ile Ser Thr Asp Tyr Ser Ser Gly 460 465 470gac tcc cag gga gcc caa ggg ggc tta tcc gat ggc ccc tac tcc aac 8062Asp Ser Gln Gly Ala Gln Gly Gly Leu Ser Asp Gly Pro Tyr Ser Asn 475 480 485cct tat gag aac agc ctt atc cca gcc gct gag cct ctg ccc ccc agc 8110Pro Tyr Glu Asn Ser Leu Ile Pro Ala Ala Glu Pro Leu Pro Pro Ser 490 495 500tat gtg gct tgc tct taggacacca ggctgcagat gatcagggat ccaatatgac 8165Tyr Val Ala Cys Ser 505tcagagaacc agtgcagact caagacttat ggaacaggga tggcgaggcc tctctcagga 8225gcaggggcat tgctgatttt gtctgcccaa tccatcctgc tcaggaaacc acaaccttgc 8285agtattttta aatatgtata gttttttttt gtatctatat atatatatac acatatgtat 8345gtaagttttt ctaccatgat ttctacaaac accctttaag tcccatcttc ccctgggcat 8405aggccatagg gatagaagtt aaagttcttg agcttattca gaagctggat ctgcaatctg 8465aatgctactc ataacataac aaaatagtat gttaaacagc tcttaaatct tactggctta 8525ccacattaaa tgatttctct ctcctaactc agctcaaatg ggcagccatc catggatgag 8585tcagaggttc agactcttcc agtctgtagc tctaccttct cttagggtac ttagatggat 8645cc 86472508PRTHomo sapiens 2Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300Gln Leu Trp Leu Tyr Gln Asn Asp Gly Cys Leu Trp Trp Ser Pro Cys305 310 315 320Thr Pro Phe Thr Glu Asp Pro Pro Ala Ser Leu Glu Val Leu Ser Glu 325 330 335Arg Cys Trp Gly Thr Met Gln Ala Val Glu Pro Gly Thr Asp Asp Glu 340 345 350Gly Pro Leu Leu Glu Pro Val Gly Ser Glu His Ala Gln Asp Thr Tyr 355 360 365Leu Val Leu Asp Lys Trp Leu Leu Pro Arg Asn Pro Pro Ser Glu Asp 370 375 380Leu Pro Gly Pro Gly Gly Ser Val Asp Ile Val Ala Met Asp Glu Gly385 390 395 400Ser Glu Ala Ser Ser Cys Ser Ser Ala Leu Ala Ser Lys Pro Ser Pro

405 410 415Glu Gly Ala Ser Ala Ala Ser Phe Glu Tyr Thr Ile Leu Asp Pro Ser 420 425 430Ser Gln Leu Leu Arg Pro Trp Thr Leu Cys Pro Glu Leu Pro Pro Thr 435 440 445Pro Pro His Leu Lys Tyr Leu Tyr Leu Val Val Ser Asp Ser Gly Ile 450 455 460Ser Thr Asp Tyr Ser Ser Gly Asp Ser Gln Gly Ala Gln Gly Gly Leu465 470 475 480Ser Asp Gly Pro Tyr Ser Asn Pro Tyr Glu Asn Ser Leu Ile Pro Ala 485 490 495Ala Glu Pro Leu Pro Pro Ser Tyr Val Ala Cys Ser 500 50531865DNAHomo sapiensmisc_feature(1)..(1865)Mature full-length EpoR mRNA 3acttagaggc gcctggtcgg gaagggcctg gtcagctgcg tccggcggag gcagctgctg 60acccagctgt ggactgtgcc gggggtgggg gacggagggg caggagccct gggctccccg 120tggcgggggc tgtatcatgg accacctcgg ggcgtccctc tggccccagg tcggctccct 180ttgtctcctg ctcgctgggg ccgcctgggc gcccccgcct aacctcccgg accccaagtt 240cgagagcaaa gcggccttgc tggcggcccg ggggcccgaa gagcttctgt gcttcaccga 300gcggttggag gacttggtgt gtttctggga ggaagcggcg agcgctgggg tgggcccggg 360caactacagc ttctcctacc agctcgagga tgagccatgg aagctgtgtc gcctgcacca 420ggctcccacg gctcgtggtg cggtgcgctt ctggtgttcg ctgcctacag ccgacacgtc 480gagcttcgtg cccctagagt tgcgcgtcac agcagcctcc ggcgctccgc gatatcaccg 540tgtcatccac atcaatgaag tagtgctcct agacgccccc gtggggctgg tggcgcggtt 600ggctgacgag agcggccacg tagtgttgcg ctggctcccg ccgcctgaga cacccatgac 660gtctcacatc cgctacgagg tggacgtctc ggccggcaac ggcgcaggga gcgtacagag 720ggtggagatc ctggagggcc gcaccgagtg tgtgctgagc aacctgcggg gccggacgcg 780ctacaccttc gccgtccgcg cgcgtatggc tgagccgagc ttcggcggct tctggagcgc 840ctggtcggag cctgtgtcgc tgctgacgcc tagcgacctg gaccccctca tcctgacgct 900ctccctcatc ctcgtggtca tcctggtgct gctgaccgtg ctcgcgctgc tctcccaccg 960ccgggctctg aagcagaaga tctggcctgg catcccgagc ccagagagcg agtttgaagg 1020cctcttcacc acccacaagg gtaacttcca gctgtggctg taccagaatg atggctgcct 1080gtggtggagc ccctgcaccc ccttcacgga ggacccacct gcttccctgg aagtcctctc 1140agagcgctgc tgggggacga tgcaggcagt ggagccgggg acagatgatg agggccccct 1200gctggagcca gtgggcagtg agcatgccca ggatacctat ctggtgctgg acaaatggtt 1260gctgccccgg aacccgccca gtgaggacct cccagggcct ggtggcagtg tggacatagt 1320ggccatggat gaaggctcag aagcatcctc ctgctcatct gctttggcct cgaagcccag 1380cccagaggga gcctctgctg ccagctttga gtacactatc ctggacccca gctcccagct 1440cttgcgtcca tggacactgt gccctgagct gccccctacc ccaccccacc taaagtacct 1500gtaccttgtg gtatctgact ctggcatctc aactgactac agctcagggg actcccaggg 1560agcccaaggg ggcttatccg atggccccta ctccaaccct tatgagaaca gccttatccc 1620agccgctgag cctctgcccc ccagctatgt ggcttgctct taggacacca ggctgcagat 1680gatcagggat ccaatatgac tcagagaacc agtgcagact caagacttat ggaacaggga 1740tggcgaggcc tctctcagga gcaggggcat tgctgatttt gtctgcccaa tccatcctgc 1800tcaggaaacc acaaccttgc agtattttta aatatgtata gtttttttat atgtatagtt 1860ttttt 18654858DNAHomo sapiensCDS(1)..(855)EpoR Isoform 1, intron 6 insert 4atg gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt 48Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15ctc ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac 96Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30ccc aag ttc gag agc aaa gcg gcc ttg ctg gcg gcc cgg ggg ccc gaa 144Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg gtg tgt ttc tgg 192Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac tac agc ttc tcc 240Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80tac cag ctc gag gat gag cca tgg aag ctg tgt cgc ctg cac cag gct 288Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95ccc acg gct cgt ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc 336Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110gac acg tcg agc ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc 384Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125ggc gct ccg cga tat cac cgt gtc atc cac atc aat gaa gta gtg ctc 432Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag agc ggc 480Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg acg tct 528His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca ggg agc 576His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190gta cag agg gtg gag atc ctg gag ggc cgc acc gag tgt gtg ctg agc 624Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc gcg cgt atg 672Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg gag cct gtg 720Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240tcg ctg ctg acg cct agc gac ctg gac ccc ctc atc ctg acg ctc tcc 768Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255ctc atc ctc gtg gtc atc ctg gtg ctg ctg acc gtg ctc gcg ctg ctc 816Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270tcc cac cgc cgg atg gtc agg gaa ggc tcc agg agg agg tga 858Ser His Arg Arg Met Val Arg Glu Gly Ser Arg Arg Arg 275 280 2855285PRTHomo sapiens 5Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270Ser His Arg Arg Met Val Arg Glu Gly Ser Arg Arg Arg 275 280 2856954DNAHomo sapiensCDS(1)..(951)EpoR Isoform 2, intron 7 insert 6atg gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt 48Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15ctc ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac 96Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30ccc aag ttc gag agc aaa gcg gcc ttg ctg gcg gcc cgg ggg ccc gaa 144Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg gtg tgt ttc tgg 192Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac tac agc ttc tcc 240Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80tac cag ctc gag gat gag cca tgg aag ctg tgt cgc ctg cac cag gct 288Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95ccc acg gct cgt ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc 336Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110gac acg tcg agc ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc 384Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125ggc gct ccg cga tat cac cgt gtc atc cac atc aat gaa gta gtg ctc 432Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag agc ggc 480Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg acg tct 528His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca ggg agc 576His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190gta cag agg gtg gag atc ctg gag ggc cgc acc gag tgt gtg ctg agc 624Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc gcg cgt atg 672Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg gag cct gtg 720Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240tcg ctg ctg acg cct agc gac ctg gac ccc ctc atc ctg acg ctc tcc 768Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255ctc atc ctc gtg gtc atc ctg gtg ctg ctg acc gtg ctc gcg ctg ctc 816Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270tcc cac cgc cgg gct ctg aag cag aag atc tgg cct ggc atc ccg agc 864Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285cca gag agc gag ttt gaa ggc ctc ttc acc acc cac aag ggt aac ttc 912Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300cag gtt ggt gct att tct tca gct gtg gct gta cca gaa tga 954Gln Val Gly Ala Ile Ser Ser Ala Val Ala Val Pro Glu305 310 3157317PRTHomo sapiens 7Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300Gln Val Gly Ala Ile Ser Ser Ala Val Ala Val Pro Glu305 310 3158987DNAHomo sapiensCDS(1)..(984)EpoR Isoform 3, intron 7 unspliced 8atg gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt 48Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15ctc ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac 96Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30ccc aag ttc gag agc aaa gcg gcc ttg ctg gcg gcc cgg ggg ccc gaa 144Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg gtg tgt ttc tgg 192Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac tac agc ttc tcc 240Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80tac cag ctc gag gat gag cca tgg aag ctg tgt cgc ctg cac cag gct 288Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95ccc acg gct cgt ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc 336Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110gac acg tcg agc ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc 384Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125ggc gct ccg cga tat cac cgt gtc atc cac atc aat gaa gta gtg ctc 432Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag agc ggc 480Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg acg tct 528His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca ggg agc 576His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190gta cag agg gtg gag atc ctg gag ggc cgc acc gag tgt gtg ctg agc 624Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc gcg cgt atg 672Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg gag cct gtg 720Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240tcg ctg ctg acg cct agc gac ctg gac ccc ctc atc ctg acg ctc tcc 768Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255ctc atc ctc gtg gtc atc ctg gtg ctg ctg acc gtg ctc gcg ctg ctc 816Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270tcc cac cgc cgg gct ctg aag cag aag atc tgg cct ggc atc ccg agc

864Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285cca gag agc gag ttt gaa ggc ctc ttc acc acc cac aag ggt aac ttc 912Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300cag gta ggt ggc ctg gtt gtc ccc tca gtg cct ggg ctt ccc tgc ttc 960Gln Val Gly Gly Leu Val Val Pro Ser Val Pro Gly Leu Pro Cys Phe305 310 315 320ttg cag cca aac tgc agg cct ctc tga 987Leu Gln Pro Asn Cys Arg Pro Leu 3259328PRTHomo sapiens 9Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Asp Leu Asp Pro Leu Ile Leu Thr Leu Ser 245 250 255Leu Ile Leu Val Val Ile Leu Val Leu Leu Thr Val Leu Ala Leu Leu 260 265 270Ser His Arg Arg Ala Leu Lys Gln Lys Ile Trp Pro Gly Ile Pro Ser 275 280 285Pro Glu Ser Glu Phe Glu Gly Leu Phe Thr Thr His Lys Gly Asn Phe 290 295 300Gln Val Gly Gly Leu Val Val Pro Ser Val Pro Gly Leu Pro Cys Phe305 310 315 320Leu Gln Pro Asn Cys Arg Pro Leu 32510804DNAHomo sapiensCDS(1)..(801)EpoR Isoform 4, intron 5 unspliced 10atg gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt 48Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15ctc ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac 96Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30ccc aag ttc gag agc aaa gcg gcc ttg ctg gcg gcc cgg ggg ccc gaa 144Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg gtg tgt ttc tgg 192Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac tac agc ttc tcc 240Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80tac cag ctc gag gat gag cca tgg aag ctg tgt cgc ctg cac cag gct 288Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95ccc acg gct cgt ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc 336Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110gac acg tcg agc ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc 384Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125ggc gct ccg cga tat cac cgt gtc atc cac atc aat gaa gta gtg ctc 432Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag agc ggc 480Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg acg tct 528His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca ggg agc 576His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190gta cag agg gtg gag atc ctg gag ggc cgc acc gag tgt gtg ctg agc 624Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc gcg cgt atg 672Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg gag cct gtg 720Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240tcg ctg ctg acg cct agc ggt gag gcc cca ggc ggg ggt gta gga gga 768Ser Leu Leu Thr Pro Ser Gly Glu Ala Pro Gly Gly Gly Val Gly Gly 245 250 255gcc agg gcg aat cac ggg gca agc cca ccg ccc tga 804Ala Arg Ala Asn His Gly Ala Ser Pro Pro Pro 260 26511267PRTHomo sapiens 11Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Gly Glu Ala Pro Gly Gly Gly Val Gly Gly 245 250 255Ala Arg Ala Asn His Gly Ala Ser Pro Pro Pro 260 26512747DNAHomo sapiensCDS(1)..(744)EpoR Isoform 5, exon 6 skipped 12atg gac cac ctc ggg gcg tcc ctc tgg ccc cag gtc ggc tcc ctt tgt 48Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15ctc ctg ctc gct ggg gcc gcc tgg gcg ccc ccg cct aac ctc ccg gac 96Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30ccc aag ttc gag agc aaa gcg gcc ttg ctg gcg gcc cgg ggg ccc gaa 144Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45gag ctt ctg tgc ttc acc gag cgg ttg gag gac ttg gtg tgt ttc tgg 192Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60gag gaa gcg gcg agc gct ggg gtg ggc ccg ggc aac tac agc ttc tcc 240Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80tac cag ctc gag gat gag cca tgg aag ctg tgt cgc ctg cac cag gct 288Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95ccc acg gct cgt ggt gcg gtg cgc ttc tgg tgt tcg ctg cct aca gcc 336Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110gac acg tcg agc ttc gtg ccc cta gag ttg cgc gtc aca gca gcc tcc 384Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125ggc gct ccg cga tat cac cgt gtc atc cac atc aat gaa gta gtg ctc 432Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140cta gac gcc ccc gtg ggg ctg gtg gcg cgg ttg gct gac gag agc ggc 480Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160cac gta gtg ttg cgc tgg ctc ccg ccg cct gag aca ccc atg acg tct 528His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175cac atc cgc tac gag gtg gac gtc tcg gcc ggc aac ggc gca ggg agc 576His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190gta cag agg gtg gag atc ctg gag ggc cgc acc gag tgt gtg ctg agc 624Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205aac ctg cgg ggc cgg acg cgc tac acc ttc gcc gtc cgc gcg cgt atg 672Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220gct gag ccg agc ttc ggc ggc ttc tgg agc gcc tgg tcg gag cct gtg 720Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240tcg ctg ctg acg cct agc ggg ctc tga 747Ser Leu Leu Thr Pro Ser Gly Leu 24513248PRTHomo sapiens 13Met Asp His Leu Gly Ala Ser Leu Trp Pro Gln Val Gly Ser Leu Cys1 5 10 15Leu Leu Leu Ala Gly Ala Ala Trp Ala Pro Pro Pro Asn Leu Pro Asp 20 25 30Pro Lys Phe Glu Ser Lys Ala Ala Leu Leu Ala Ala Arg Gly Pro Glu 35 40 45Glu Leu Leu Cys Phe Thr Glu Arg Leu Glu Asp Leu Val Cys Phe Trp 50 55 60Glu Glu Ala Ala Ser Ala Gly Val Gly Pro Gly Asn Tyr Ser Phe Ser65 70 75 80Tyr Gln Leu Glu Asp Glu Pro Trp Lys Leu Cys Arg Leu His Gln Ala 85 90 95Pro Thr Ala Arg Gly Ala Val Arg Phe Trp Cys Ser Leu Pro Thr Ala 100 105 110Asp Thr Ser Ser Phe Val Pro Leu Glu Leu Arg Val Thr Ala Ala Ser 115 120 125Gly Ala Pro Arg Tyr His Arg Val Ile His Ile Asn Glu Val Val Leu 130 135 140Leu Asp Ala Pro Val Gly Leu Val Ala Arg Leu Ala Asp Glu Ser Gly145 150 155 160His Val Val Leu Arg Trp Leu Pro Pro Pro Glu Thr Pro Met Thr Ser 165 170 175His Ile Arg Tyr Glu Val Asp Val Ser Ala Gly Asn Gly Ala Gly Ser 180 185 190Val Gln Arg Val Glu Ile Leu Glu Gly Arg Thr Glu Cys Val Leu Ser 195 200 205Asn Leu Arg Gly Arg Thr Arg Tyr Thr Phe Ala Val Arg Ala Arg Met 210 215 220Ala Glu Pro Ser Phe Gly Gly Phe Trp Ser Ala Trp Ser Glu Pro Val225 230 235 240Ser Leu Leu Thr Pro Ser Gly Leu 2451422DNAArtificial sequenceSynthetic oligonucleotide 14tcaagcggct gcttccttcc aa 221524DNAArtificial sequenceSynthetic oligonucleotide 15gcagggagcg tacagagggt ggag 241624DNAArtificial sequenceSynthetic oligonucleotide 16gaagaaatag caccaacctg gaag 241722DNAArtificial sequenceSynthetic oligonucleotide 17ctgacgccta gcgacctgga cc 221821DNAArtificial sequenceSynthetic oligonucleotide 18gcagtttggc tgcaagaagc a 211920DNAArtificial sequenceSynthetic oligonucleotide 19ggagccaggg cgaatcacgg 202020DNAArtificial sequenceSynthetic oligonucleotide 20gccttcaaac tcgctctctg 202121DNAArtificial sequenceSynthetic oligonucleotide 21gcttcagagc ccgctaggcg t 21

Claims

1. An isolated nucleic acid selected from the group consisting ofa nucleic acid encoding erythropoietin receptor isoform 3 and having the sequence given herein as SEQ ID NO: 8;a nucleic acid encoding erythropoietin receptor isoform 4 and having the sequence given herein as SEQ ID NO: 10;a nucleic acid encoding erythropoietin receptor isoform 5 and having the sequence given herein as SEQ ID NO: 12;a nucleic acid that encodes the opposite strand of a nucleic acid as set forth above.

2-3. (canceled)

4. The nucleic acid according to claim 1 encoding erythropoietin receptor isoform 3 and having the sequence given herein as SEQ ID NO: 8.

5. The nucleic acid according to claim 1 encoding erythropoietin receptor isoform 4 and having the sequence given herein as SEQ ID NO: 10.

6. The nucleic acid according to claim 1 encoding erythropoietin receptor isoform 5 and having the sequence given herein as SEQ ID NO: 12.

7. The nucleic acid according to claim 1, wherein said nucleic acid is an RNA.

8. A recombinant nucleic acid comprising a promoter operatively associated with a nucleic acid according to claim 1.

9. A host cell containing a recombinant nucleic acid according to claim 8 and which expresses the encoded erythropoetin receptor isoform.

10-16. (canceled)

17. A method of screening a subject for cancer, comprising:detecting the presence or absence of a nucleic acid according to claim 1 in said subject,the presence of such a nucleic acid indicating said subject is afflicted with or at risk of developing cancer.

18. The method according to claim 17, wherein said cancer is breast, cervix, colon, lung, ovarian or prostate cancer.

19. The method according to claim 17, wherein said detecting step is carried out by collecting a biological sample from said subject, and then detecting the presence or absence of said nucleic acid in said biological sample.

20. A method of screening a subject for cancer, comprising:detecting the presence or absence of a protein encoded by a nucleic acid according to claim 1 in said subject,the presence of such a protein indicating said subject is afflicted with or at risk of developing cancer.

21. The method according to claim 20, wherein said cancer is breast, cervix, colon, lung, ovarian or prostate cancer.

22. The method according to claim 20, wherein said detecting step is carried out by collecting a biological sample from said subject, and then detecting the presence or absence of said protein in said biological sample.

23. The method according to claim 20, wherein said detecting step is carried out by immunoassay.

24. The method according to claim 20, wherein said detecting step is carried out by detecting nucleic acid that encodes said protein.