WO1994007916A1 - Recepteur d'hormone steroide humain neri - Google Patents

Recepteur d'hormone steroide humain neri

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Publication number
WO1994007916A1
WO1994007916A1 PCT/US1993/009165 US9309165W WO9407916A1 WO 1994007916 A1 WO1994007916 A1 WO 1994007916A1 US 9309165 W US9309165 W US 9309165W WO 9407916 A1 WO9407916 A1 WO 9407916A1
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WIPO (PCT)
Prior art keywords
ser
leu
gin
glu
pro
Prior art date
Application number
PCT/US1993/009165
Other languages
English (en)
Inventor
Azriel Schmidt
Gideon A. Rodan
Su Jane Rutledge
Robert L. Vogel
Original Assignee
Merck & Co., Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Merck & Co., Inc. filed Critical Merck & Co., Inc.
Priority to AU51651/93A priority Critical patent/AU5165193A/en
Publication of WO1994007916A1 publication Critical patent/WO1994007916A1/fr

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Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/721Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor

Definitions

  • the present invention relates generally to ligand-responsive regulatory proteins and genes encoding them.
  • a novel recombinant human steroid hormone receptor hereinafter identified as NERI
  • NERI human steroid hormone receptor
  • expression systems including a COS stable expression system
  • assay using the COS expression system is also disclosed.
  • the invention relates to a method for identifying functional ligands of the NERI receptor.
  • Retinoids, steroid and thyroid hormones and possibly other molecules produce their biological effects by binding to proteins of the steroid receptor superfamily. These receptors interact with specific DNA sequences and modulate gene expression (for reviews see JM Berg, Cell 57:1065-1068 (1989); RM Evans, Science 240:899-895 (1988); M Beato, Cell 56:335-344 (1989)). Sequence analysis and functional studies of these receptors revealed two important regions which exhibit a high degree of amino acid residue conservation.
  • the highest level of similarity among the receptors is found in a region which contains nine cystein residues that bind zinc atoms to form two "zinc fingers," which interact with the cognate steroid response elements of DNA (J Miller, et al., EMBO J 4:1609-1614 (1985); RM Evans, Cell 52:1-3 (1988)).
  • the second region which is less conserved, is the ligand binding domain, responsible for the interaction with the hormone (J Carlstedt-Duke, et al, Proc Natl Acad Sci USA 79:4260-4264 (1982). J. Carlstedt-Duke, et ah, Proc Natl Acad Sci USA 84:4437- 4440 (1987)).
  • hERl and hER2 have been cloned by low stringency hybridization of cDNA libraries with a DNA probe coding for the DNA binding domain of the estrogen receptor (V Giguere, et al., Nature 331:91-94 (1988)). Similar approaches have led to the discovery of the retinoic acid receptors and the peroxisome proliferator activator receptor (PPAR)(I Issemann, et al., Nature 347:645-650 (1990); DJ Mangelsdorf, et al, Nature 345:224-229 (1990)). Recently, three novel members of the Xenopus nuclear hormone receptor superfamily have been disclosed (C Dreyer, Cell 68:879-887 (1992)).
  • Fig. 1 shows the cDNA sequence of the human NER-1 receptor (Seq. ID No. 1) and the associated expression protein (Seq. ID No. 2).
  • the circled P indicates the amino acid proline and the boxed area represents the binding area of the protein to other DNA.
  • Fig. 2 illustrates in A, the three probes used for NER-1, ES 11 (Seq. ID No. 3), ES 12 (Seq. ID No. 4), and antisense ES 15 (Seq. ID No. 5).
  • B illustrates the binding region between the ES 12 probe and the NER-1 cDNA.
  • One embodiment of the invention concerns human steroid hormone receptor NERI, said receptor being free of other human receptor proteins. In one class this embodiment concerns human steroid hormone receptor NERI, said receptor being free of other human proteins.
  • this embodiment concerns human steroid hormone receptor NERI from human cells such as osteosarcoma, said receptor being free of other human proteins.
  • this embodiment concerns a protein comprising the following 461 amino acid sequence (SEQ ID NO:2:) depicted from the amino to the carboxy terminus:
  • a second embodiment concerns a DNA sequence encoding human steroid hormone receptor NERI complementary DNA, said DNA, said sequence being free of other human DNA sequences.
  • the invention also includes alternative base sequences wherein a codon (or codons) are replaced with another codon, such that the amino acid sequence translated by the DNA sequence remains unchanged.
  • a sequence bearing one or more such replaced codons will be defined as a degenerate variation.
  • mutations exchange of individual amino acids which one of skill in the art would expect to have no effect on functionality, such as valine for leucine, arginine for lysine and asparagine for glutamine.
  • One class of the second embodiment of the invention concerns the following nucleotide sequence (SEQ ID NO:l:) of complementary DNA depicted from the 5' to the 3' terminus:
  • CAAGAAGTGG CGAAGTTACC TTTGAGGGTA TTTGAGTAGC GGCGGTGTGT CAGGGGCTAA 60
  • CACCATGTCC TCTCCTACCA CGAGTTCCCT GGATACCCCC CTGCCTGGAA ATGGCCCCCC 300
  • a third embodiment of this invention concerns systems for expressing all or part of the human steroid hormone receptor NERI.
  • One class of this third embodiment of the invention comprises: An expression construct, such as a plasmid which comprises: a) an expression vector, such as PJ3NERI, and b) a base sequence encoding human steroid hormone receptor NERI protein.
  • the steroid hormone receptor NERI comprises the nucleotide sequence (SEQ ID NO:l:) of complementary DNA as shown above.
  • a second class of this third embodiment of the invention concerns a system for the transient expression of human steroid hormone receptor NERI in a suitable host cell, such as a monkey kidney cell line (COS), the system comprised of a vector which expresses human steroid hormone receptor NERI cDNA.
  • COS monkey kidney cell line
  • Suitable cell lines derived from various species include, but are not limited to, cell lines of human, bovine, porcine, monkey, and rodent origin, or from yeast and bacterial strains.
  • a fourth embodiment of the invention concerns a method of using any of the above eukaryote or prokaryote expression systems for determining the binding affinity of a test sample for steroid hormone receptor NERI.
  • a chimeric gene can be created by substituting the DNA-binding domain region in the DNA sequence encoding NERI cDNA with a DNA-binding domain region taken from a DNA sequence coding for another steroid hormone receptor protein, e.g., glucocorticoid (GR) receptor protein, thyroid receptor protein, mineral-ocorticoid receptor protein or retinoic acid receptor protein.
  • GR glucocorticoid
  • a suitable receptor-deficient host cell is transfected with: (1) the chimeric receptor gene, which is preferably carried on an expression plasmid, and (2) a reporter gene, such as the CAT gene or the firefly luciferase gene, which is also preferably carried on a plasmid.
  • the reporter gene is functionally linked to an operative hormone response element (HRE) (either wild-type or engineered) wherein the hormone response element is capable of being activated by the DNA- binding domain used to make the chimeric receptor gene.
  • HRE operative hormone response element
  • the HRE should be a wild-type, an engineered, or a synthetic GRE, i.e., one that can be activated by the operative portion of the DNA-binding region of a GR receptor protein.
  • the transfected host cell is challenged with a test sample which contains one or more ligand(s) which can potentially bind with the ligand-binding domain region of the chimeric protein coded for by the chimeric gene.
  • the fourth embodiment further concerns a method for determining the affinity of a test sample for activation of a steroid hormone receptor NERI, the method comprising:
  • step (i) the chimeric gene from step (a), and (ii) a reporter gene functionally linked to an operative hormone response element wherein the hormone response element is capable of being activated by the DNA-binding domain region of the receptor protein encoded by the chimeric gene of step (a);
  • step (c) challenging the transfected host cell from step (b) with the test sample to be evaluated for ligand-binding activity with the chimeric receptor protein encoded by the chimeric gene of step (a);
  • COS monkey kidney cell line
  • plasmid the plasmid comprising:
  • the aforementioned fourth embodiment is further useful for identifying compounds which may be peroxisome proliferators and, hence, are potentially hepatocarcinogens.
  • This embodiment is also useful in identifying ligands for new hormone systems which regulate bodily function.
  • the present invention describes methods to isolate the human steroid hormone receptor NERI complementary DNA (cDNA) without prior knowledge of its protein sequence or gene sequence.
  • PCR Polymerase chain reaction
  • the complete sequence of the human steroid hormone receptor NERI cDNA was determined, and its encoded protein sequence was deduced. Among other things, such sequence information is useful in the process of developing novel steroid hormone agonists and antagonists.
  • An expression system was used to express the cloned human steroid hormone receptor NERI cDNA.
  • the COS (a monkey kidney cell line) expression system can be used to measure the ligand binding properties of human steroid hormone receptor NERI.
  • Assay protocols use the heterologously expressed human steroid hormone receptor NERI for determination of the activation of steroid hormone receptor NERI by antagonists.
  • the present invention generally relates to a new member of the steroid hormone receptor superfamily.
  • the amino acid sequence deduced from the DNA sequence shows the characteristic features of both the DNA and the ligand binding domains of this family of receptors.
  • Sequence analysis predicted a protein of 461 amino acids which includes the conserved amino acid residues characteristic of the DNA and ligand-binding domains of nuclear receptors.
  • Ner-I a new member of the steroid receptor-like gene family which was isolated from a human bone cell cDNA library.
  • Ner-I codes for a polypeptide of 461 amino acids which contains the conserved sequences of the DNA and ligand binding domains of typical steroid receptors. The best homology is shared with the different retinoic acid receptors: ⁇ , ⁇ & ⁇ , 55% at the DNA ⁇ , ⁇ binding domain and 38-40% at the ligand binding domain.
  • a single transcript of 2.3kb was detected in all cells and tissues tested. We tested the potential of these constructs to mediate ligand dependent transcription activation of reporter genes. To date, no specific ligand for this receptor was identified but it is reasonably believed that binding will occur with a member of a retinoic acid receptor family.
  • the nuclear receptor-gene family is expanding in size, as new members are constantly identified.
  • This gene named Ner-I, codes for a polypeptide of 461 amino acids and contains the conserved sequences typical of both the DNA and the ligand binding domains.
  • the amino terminal of the predicted protein contains a high number of proline and serine residues which might introduce a highly stabilized and complexed secondary structure.
  • a high number of proline residues was also found in other nuclear receptor and other molecules with transcriptional activity such as CTf/Nl, fos, jun. p53, OCT-2 and SRF (Mitchell & Tjian, Science, 245, pp. 371-378 (1989); Meimod et al 1989).
  • the size of the deduced protein and the spatial distribution of the different domains resemble the arrangement found in the thyroid, vitamin D and retinoic acid-receptor subgroup (Lazar et al.. Proc Natl. Acad. Sci. 86, pp. 7771-7774, 1989).
  • the sequence homology at the predicted ligand binding domain ranges between 33-40% identity with the members of this subgroup, while homologies lower than 25% were measured when the ligand binding domain was compared to the corresponding domain of the steroid receptor subgroup which includes the estrogen, glucocorticoid, androgen and progesterone.
  • the highest homology of the ligand binding domain was the retinoic acid receptors.
  • the mRNA for Ner-I is widely distributed in different tissues and in all the tested cell lines.
  • steroid hormone receptor superfamily refers to the class of related receptors comprised of glucocorticoid, mineralocorticoid, progesterone, estrogen, estrogen-related, vitamin D3, thyroid, v-erb-A, retinoic acid and E75 (Drosophilia) receptors.
  • steroid hormone receptor refers to members within the steroid hormone receptor superfamily.
  • ligand means an inducer, such as a hormone or growth substance. Inside a cell the ligand binds to a receptor protein, thereby creating a ligand-receptor complex, which in turn can bind to an appropriate hormone response element. Single ligands may have multiple receptors.
  • expression construct refers to a plasmid or vector comprising a transcriptional unit comprising an assembly of (1) a genetic element or elements having a regulatory role in gene expression, for example, promoters or enhancers, (2) a structural or coding sequence which is transcribed into mRNA and translated into protein, and (3) appropriate transcription initiation and termination sequences.
  • Recombinant expression system means a combination of an expression construct and a suitable host microorganism.
  • Degenerate DNA primers were designed to recognize the consensus sequences of the DNA and ligand binding domains of a typical nuclear receptor.
  • the 5' primer ESl l (Seq. ID. No. 3) was degenerate oligomer 5' TGTGAGGGCTGCAA(G/A)G(C/G)C, based on the conserved amino acids CEGCKA(G) of the DNA binding domain.
  • a second 5' primer, ESI 2, (Seq. ID. No. 4)
  • TGTGAGGGCTGCAA(G/A)G(C/G)CTTCTTC contains six additional nucleodites at its 3'-end corresponding to two conserved phenyalanine residues following the CEGCKA(G) sequence.
  • the antisense primer, ES15 (Seq. ID. No. 5)
  • AA(G)A(C,T,G)CCA(C,T,G)GGIA ⁇ IIC(T)TTT(A,G,C)GC(G)TT was designed to complement the semiconserved aminoacid sequence FAKxxPGF of the ligand binding domain of a typical receptor.
  • the nucleotides corresponding to the nonconserved aminoacids (XX) were substituted with inosine (I) residues.
  • PCR polymerase chain reaction
  • a random primed cDNA library was prepared from 2 ⁇ g total RNA isolated from the osteosarcoma SAOS-2/B10 cells by the Moloney reverse transcriptase enzyme RTH according to the manufacturer recommendations (Bethesda Research Laboratories).
  • the cDNA reaction (25 ⁇ l) was diluted into 300 ml water and heat denatured at 95°C for 5 minutes and quickly chilled on ice.
  • the cDNA (2.5 ⁇ l) and the first primer pair, ESl l and ES15 (0.5 ⁇ M each) were employed in the amplification reaction with the amplitaq kit and the DNA thermal cycler (Perkin-Elmer-Cetus).
  • Primer ESll has the following sequence (SEQ ID NO:3:):
  • R represents A or G
  • S represents C or G; and Primer ES15 has the following sequence (SEQ ID NO:5:):
  • N (at the 11, 14 & 26 positions) represents A or C or G or T; N (at the 17, 19, 20, 21 & 22 positions) represent inosine; R represents A or G; S represents C or G; and Y represents C or T.
  • the following amplification cycles were conducted: denaturation at 94°C, 1.5 minutes; annealing at 65°C, 3 minutes; extension at 72°C, 5 minutes for 3 cycles; denaturing at 94°C, 1 minute; annealing at 60°C, 3 minutes; extension at 72°C, 5 minutes for 15 cycles; and denaturing at 94°C, 1 minute; annealing at 57°C, 3 minutes; extension at 72°C, 5 minutes for 20 cycles.
  • Primer ES12 has the following sequence (SEQ ID NO:4:):
  • S represents C or G.
  • the second round of amplification was performed with the same program used for the first amplification cycles.
  • the amplification products were separated on 5% polyacrylamide gel and stained by ethidium bromide.
  • the DNA products were isolated from the gel, phosphorylated by T4 polynucleotide kinase and cloned into PUC 18 vector by blunt end ligation. Clones were identified by digestion of plasmid DNA with PvuII enzyme.
  • the DNA insert was analyzed by double-stranded DNA sequencing by the dideoxy termination method using sequenase enzyme kit (United States Biochemicals).
  • This amplification produced two major DNA fragments of 270 bp and 320, respectively.
  • Single stranded randomly primed cDNA was prepared with the Mo-MLV reverse transcriptase (BRL) from RNA isolated from Saos-2 B10, a human osteosarcoma cell line (Rodan et al., Cancer Research, 47], pp. 4961-4966, 1987; Endocrinol, 122, pp. 219-227, (1989).
  • the cDNA reaction (25 ⁇ l) was diluted into 300 ⁇ l water and heat denaturated at 95°C for 5 minutes and quickly chilled on ice.
  • the amplitaq kit and the DNA thermal cycler Perkin Elmer, Cetus.
  • amplification reaction buffer containing a second set of primers: a partially nested oligomer ESI 2 and the same 3'-end primer, ESI 5 (0.5 ⁇ M each).
  • the second round of amplification was performed with the same program used for the first amplification cycles.
  • the amplified fragments were separated by electrophoresis on 5% polyacrylamide gel, cloned into plasmids and sequenced.
  • the DNA fragments were then used to screen a lamda gtl 1 cDNA library of Saos- 2/B10 cells. Positive clones were isolated and sequenced in a bidirectional way by the sequence kit (United States Biochemicals).
  • a human oligo-dT cDNA library was constructed RNA isolated from osteosarcoma SAOS-2/B10 cells using the Lambda Librarian cloning kit (Invitrogen Corp.). Several positive clones were identified by plaque screening with the [32p] labeled DNA probe of the cloned amplified product (NERI). The hybridization conditions were as described by A Schmidt, et al, J Biol Chem 259:7411-7415 (1984). The cDNA inserts were cloned into EcoRI site of the cloning vector PUC18. The complete DNA sequence of both strands was determined by the dideoxy sequencing method using a series of oligonucleotides synthesized as the DNA sequence data became available.
  • NERI cloned amplified product
  • the fragments from PCR amplification were cloned into plasmids and sequenced.
  • the amplified cDNA fragment of 270 bp NERI was used for the screening of a human osteosarcoma SAOS- 2/B10 cells cDNA library. All the highly positive clones were identical and matched the sequence for the amplified NERI DNA fragment.
  • RNA from various tissues or the listed cell lines were prepared by using guanidine thiocyanate or by the guanidine hydrochloride method (GGA Nemeth, et aL, Anal Biochem 183:301- 304 (1989); JM Chirgwin, et al., Biochemistry. 18:5294-5299 (1979)). RNA samples were analyzed by formaldehyde agarose gel electrophoresis as described by (KM Rosen, et al., Focus 12:23-24 (1990)).
  • the amplification of the cDNA prepared from the RNA of Saos-2/B10 osteoblastic cell line with the ESl l and ESI 5 primers yielded multiple fragments after 40 rounds of amplification.
  • Five percent of the first amplification reaction were subjected to additional 30 rounds of amplication with ESI 2 and ESI 5 oligomers.
  • Primer ESI 2 that replaces ESI 1 is six nucleodites longer and codes for two conserved phenylalanine residues at the 3'-end, thus introduces an additional level of specificity to the amplification reaction.
  • the second amplification step resulted in the elimination of all but two DNA fragments.
  • Sequencing of the Ner-I clone revealed a long open reading frame coding for a polypeptide of 461 amino acids.
  • the deduced protein resembles in its structure a typical steroid-like receptor.
  • a putative "double zinc finger" structure which can serve as the DNA binding domain was identified.
  • Amino acid sequences that characterize the ligand binding domain were located toward the carboxy terminus of the protein and were spaced like in the thyroid or retinoic acid receptors. Comparing the sequence of the deduced protein with other known receptor sequences revealed that the DNA binding domain shared 50-56% homology with all the steroid-like receptors.
  • Highest scores at this domain were: 56% for the estrogen receptor, 55% for the retinoic acid gamma receptor and mineralcorticoid receptor and 54% for retinoic acid A and glucocorticoid receptors.
  • the ligand binding domain which is less conserved showed highest homology levels of 38-40% with the 3 types of retinoic acid receptors, RAR ⁇ , RAR ⁇ and RAR ⁇ 38% with vitamin D receptor and 33% with thyroid hormone receptor.
  • the homology to the ligand binding domains of estrogen, androgen, glucocorticoid and mineralocorticoid at this domain was significantly lower.
  • the RXR retinoic acid receptor type X showed an intermediate value of 28% homology at this domain.
  • Ner-I amino acids 1-87
  • RNA from the osteoblastic Saos-2/B10 cells with the Ner-I labeled DNA probe revealed a single transcript of approximate 2.3 kb. Similar RNA transcripts were detected in all cell lines tested. No apparent variations in size of the mRNA molecules could be observed between RNAs isolated from different species. Tissue distribution of the Ner-I gene expression was examined by Northern hybridization. Ner-I RNA transcripts were detected in all the rat tissues which were tested. Similar results were obtained with RNA isolated from tissues of adult baboons.
  • This recombinant receptor was employed in ligand transcription experiments using the pERE-BLCAT plasmid as a reporter gene (Lukow and Schultz, Nuc. Acid Res. 15, pp. 5490-5491, (1987) or MMTV-luciferase reporter plasmid.
  • a reporter gene Likow and Schultz, Nuc. Acid Res. 15, pp. 5490-5491, (1987) or MMTV-luciferase reporter plasmid.
  • a steroid hormone exists for which NER-I binds to and associated with biological activity at concentrations of 1-10 micromolar.
  • Northern analysis with a NERI cDNA probe revealed that NERI receptor mRNA is expressed as a 2.3 Kb transcript in the human osteosar
  • MOLECULE TYPE DNA (genomic)
  • HYPOTHETICAL NO
  • ANTI -SENSE NO

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Abstract

Un nouveau membre de la superfamille des récepteurs d'hormones stéroïdes (identifié sous le nom de NERI) est décrit et a été préparé par clonage d'ADNc à partir d'une librairie de cellules d'ostéosarcome humaines SAOS-2/B10. L'invention décrit également la séquence complète de l'ADN complémentaire NERI humain (No. de la Séq. 1); des systèmes d'expression, y compris un système d'expression stable COS; la protéine exprimée (No. de Séq. 2) et un dosage utilisant le système d'expression COS. NERI peut être utilisé dans un dosage afin d'identifier et évaluer les entités chimiques qui se lient à ce récepteur.
PCT/US1993/009165 1992-10-07 1993-09-27 Recepteur d'hormone steroide humain neri WO1994007916A1 (fr)

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AU51651/93A AU5165193A (en) 1992-10-07 1993-09-27 Human steroid hormone receptor neri

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US95813792A 1992-10-07 1992-10-07
US958,137 1992-10-07

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WO1994007916A1 true WO1994007916A1 (fr) 1994-04-14

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WO1995013373A1 (fr) * 1993-11-10 1995-05-18 Arch Development Corporation Recepteur nucleaire ubiquiste: compositions et procedes
WO1996009324A1 (fr) * 1994-09-21 1996-03-28 Cancer Institute Proteine ecdn et adn codant cette proteine
WO1996013519A1 (fr) * 1994-10-27 1996-05-09 Merck & Co., Inc. Activation d'un recepteur de steroïde humain
WO1998001552A2 (fr) * 1996-07-09 1998-01-15 Genetics Institute, Inc. Proteines secretees et polynucleotides codant lesdites proteines
WO1998001554A2 (fr) * 1996-07-09 1998-01-15 Genetics Institute, Inc. Proteines secretees et polynucleotides codant lesdites proteines
WO1998001469A2 (fr) * 1996-06-19 1998-01-15 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998004695A1 (fr) * 1996-07-26 1998-02-05 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998004696A1 (fr) * 1996-07-26 1998-02-05 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
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WO1998004694A2 (fr) * 1996-07-26 1998-02-05 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998005781A1 (fr) * 1996-08-02 1998-02-12 Genetics Institute, Inc. Proteines secretees et polynucleotides codant pour elles
WO1998005776A2 (fr) * 1996-08-02 1998-02-12 Genetics Institute, Inc. Proteines secretees et polynucleotides codant ces proteines
WO1998007856A1 (fr) * 1996-08-23 1998-02-26 Genetics Institute, Inc. Proteines secretees et polynucleotides codant pour ces proteines
WO1998007855A2 (fr) * 1996-08-23 1998-02-26 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998007859A2 (fr) * 1996-08-23 1998-02-26 Genetics Institute, Inc. Proteines secretees et polynucleotides codant lesdites proteines
WO1998007853A1 (fr) * 1996-08-23 1998-02-26 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
US5728548A (en) * 1995-06-29 1998-03-17 Genetics Institute, Inc. Retinoid receptor-1 (RR1) and DNA encoding RR1
WO1998014575A1 (fr) * 1996-10-04 1998-04-09 Genetics Institute, Inc. Proteines secretees et polynucleotides codant pour celles-ci
WO1998014576A2 (fr) * 1996-10-04 1998-04-09 Genetics Institute, Inc. Proteines secretees et polynucleotides codant pour celles-ci
WO1998014470A2 (fr) * 1996-10-04 1998-04-09 Genetics Institute, Inc. Proteines secretees
WO1998017687A2 (fr) * 1996-10-25 1998-04-30 Genetics Institute, Inc. Proteines secretees et polynucleotides codant ces proteines
WO1998020130A2 (fr) * 1996-11-01 1998-05-14 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998020125A1 (fr) * 1996-11-06 1998-05-14 Genetics Institute, Inc. Proteines secretees et polynucleotides qui les codent
WO1998021332A2 (fr) * 1996-11-15 1998-05-22 Genetics Institute, Inc. Proteines secretes et polynucleotides les codant
WO1998024905A2 (fr) * 1996-12-06 1998-06-11 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998030696A2 (fr) * 1997-01-13 1998-07-16 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998030695A2 (fr) * 1997-01-09 1998-07-16 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998031802A2 (fr) * 1997-01-21 1998-07-23 Genetics Institute, Inc. Proteines secretees et polynucleotides codant ces proteines
WO1998032853A2 (fr) * 1997-01-24 1998-07-30 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998033916A2 (fr) * 1997-01-31 1998-08-06 Genetics Institute, Inc. Proteines secretees et polynucleotides codant celles-ci
WO1998037094A2 (fr) * 1997-02-24 1998-08-27 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
WO1998040486A2 (fr) * 1997-03-14 1998-09-17 Genetics Institute, Inc. Proteines secretees et polynucleotides les codant
US5945302A (en) * 1996-04-05 1999-08-31 Genetics Institute, Inc. Polynucleotides encoding secreted proteins
WO2005101007A1 (fr) * 2004-04-15 2005-10-27 Bayer Healthcare Ag Diagnostic et traitement de maladies associees au recepteur beta du foie x (lxrb)

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Cited By (64)

* Cited by examiner, † Cited by third party
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