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. 1993 Oct 1;90(19):8967–8971. doi: 10.1073/pnas.90.19.8967

Expression cloning of a human corticotropin-releasing-factor receptor.

R Chen 1, K A Lewis 1, M H Perrin 1, W W Vale 1
PMCID: PMC47482  PMID: 7692441

Abstract

Corticotropin-releasing factor (CRF) is the principal neuroregulator of the hypothalamic-pituitary-adrenocortical axis and plays an important role in coordinating the endocrine, autonomic, and behavioral responses to stress and immune challenge. We report here the cloning of a cDNA coding for a CRF receptor from a human corticotropic tumor library. The cloned cDNA encodes a 415-amino acid protein comprising seven putative membrane-spanning domains and is structurally related to the calcitonin/vasoactive intestinal peptide/growth hormone-releasing hormone subfamily of G protein-coupled receptors. The receptor expressed in COS cells binds rat/human CRF with high affinity (Kd = 3.3 +/- 0.45 nM) and specificity and is functionally coupled to adenylate cyclase. The CRF antagonist alpha-helCRF-(9-41) inhibits the CRF-stimulated increase in intracellular cAMP. Northern blot analysis reveals that the CRF receptor is expressed in the rat pituitary and brain as well as in the mouse AtT20 corticotropic cells. We also describe an alternatively spliced form of the receptor which includes an insert of 29 amino acids in the first intracellular loop.

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Selected References

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  1. Aguilera G., Harwood J. P., Wilson J. X., Morell J., Brown J. H., Catt K. J. Mechanisms of action of corticotropin-releasing factor and other regulators of corticotropin release in rat pituitary cells. J Biol Chem. 1983 Jul 10;258(13):8039–8045. [PubMed] [Google Scholar]
  2. Behan D. P., Linton E. A., Lowry P. J. Isolation of the human plasma corticotrophin-releasing factor-binding protein. J Endocrinol. 1989 Jul;122(1):23–31. doi: 10.1677/joe.0.1220023. [DOI] [PubMed] [Google Scholar]
  3. Bilezikjian L. M., Vale W. W. Glucocorticoids inhibit corticotropin-releasing factor-induced production of adenosine 3',5'-monophosphate in cultured anterior pituitary cells. Endocrinology. 1983 Aug;113(2):657–662. doi: 10.1210/endo-113-2-657. [DOI] [PubMed] [Google Scholar]
  4. Bilezikjian L. M., Woodgett J. R., Hunter T., Vale W. W. Phorbol ester-induced down-regulation of protein kinase C abolishes vasopressin-mediated responses in rat anterior pituitary cells. Mol Endocrinol. 1987 Aug;1(8):555–560. doi: 10.1210/mend-1-8-555. [DOI] [PubMed] [Google Scholar]
  5. Crofford L. J., Sano H., Karalis K., Webster E. L., Goldmuntz E. A., Chrousos G. P., Wilder R. L. Local secretion of corticotropin-releasing hormone in the joints of Lewis rats with inflammatory arthritis. J Clin Invest. 1992 Dec;90(6):2555–2564. doi: 10.1172/JCI116150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cronin M. J., Zysk J. R., Baertschi A. J. Protein kinase C potentiates corticotropin releasing factor stimulated cyclic AMP in pituitary. Peptides. 1986 Sep-Oct;7(5):935–938. doi: 10.1016/0196-9781(86)90114-2. [DOI] [PubMed] [Google Scholar]
  7. De Souza E. B., Perrin M. H., Insel T. R., Rivier J., Vale W. W., Kuhar M. J. Corticotropin-releasing factor receptors in rat forebrain: autoradiographic identification. Science. 1984 Jun 29;224(4656):1449–1451. doi: 10.1126/science.6328656. [DOI] [PubMed] [Google Scholar]
  8. De Souza E. B., Perrin M. H., Whitehouse P. J., Rivier J., Vale W., Kuhar M. J. Corticotropin-releasing factor receptors in human pituitary gland: autoradiographic localization. Neuroendocrinology. 1985 May;40(5):419–422. doi: 10.1159/000124107. [DOI] [PubMed] [Google Scholar]
  9. De Souza E. B., Whitehouse P. J., Kuhar M. J., Price D. L., Vale W. W. Reciprocal changes in corticotropin-releasing factor (CRF)-like immunoreactivity and CRF receptors in cerebral cortex of Alzheimer's disease. Nature. 1986 Feb 13;319(6054):593–595. doi: 10.1038/319593a0. [DOI] [PubMed] [Google Scholar]
  10. DeLean A., Munson P. J., Rodbard D. Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Am J Physiol. 1978 Aug;235(2):E97–102. doi: 10.1152/ajpendo.1978.235.2.E97. [DOI] [PubMed] [Google Scholar]
  11. Dohlman H. G., Thorner J., Caron M. G., Lefkowitz R. J. Model systems for the study of seven-transmembrane-segment receptors. Annu Rev Biochem. 1991;60:653–688. doi: 10.1146/annurev.bi.60.070191.003253. [DOI] [PubMed] [Google Scholar]
  12. Gaylinn B. D., Harrison J. K., Zysk J. R., Lyons C. E., Lynch K. R., Thorner M. O. Molecular cloning and expression of a human anterior pituitary receptor for growth hormone-releasing hormone. Mol Endocrinol. 1993 Jan;7(1):77–84. doi: 10.1210/mend.7.1.7680413. [DOI] [PubMed] [Google Scholar]
  13. Grigoriadis D. E., De Souza E. B. Heterogeneity between brain and pituitary corticotropin-releasing factor receptors is due to differential glycosylation. Endocrinology. 1989 Oct;125(4):1877–1888. doi: 10.1210/endo-125-4-1877. [DOI] [PubMed] [Google Scholar]
  14. Ishihara T., Nakamura S., Kaziro Y., Takahashi T., Takahashi K., Nagata S. Molecular cloning and expression of a cDNA encoding the secretin receptor. EMBO J. 1991 Jul;10(7):1635–1641. doi: 10.1002/j.1460-2075.1991.tb07686.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ishihara T., Shigemoto R., Mori K., Takahashi K., Nagata S. Functional expression and tissue distribution of a novel receptor for vasoactive intestinal polypeptide. Neuron. 1992 Apr;8(4):811–819. doi: 10.1016/0896-6273(92)90101-i. [DOI] [PubMed] [Google Scholar]
  16. Kennelly P. J., Krebs E. G. Consensus sequences as substrate specificity determinants for protein kinases and protein phosphatases. J Biol Chem. 1991 Aug 25;266(24):15555–15558. [PubMed] [Google Scholar]
  17. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  18. Labrie F., Veilleux R., Lefevre G., Coy D. H., Sueiras-Diaz J., Schally A. V. Corticotropin-releasing factor stimulates accumulation of adenosine 3', 5'-monophosphate in rat pituitary corticotrophs. Science. 1982 May 28;216(4549):1007–1008. doi: 10.1126/science.6281886. [DOI] [PubMed] [Google Scholar]
  19. Lin C., Lin S. C., Chang C. P., Rosenfeld M. G. Pit-1-dependent expression of the receptor for growth hormone releasing factor mediates pituitary cell growth. Nature. 1992 Dec 24;360(6406):765–768. doi: 10.1038/360765a0. [DOI] [PubMed] [Google Scholar]
  20. Lin H. Y., Harris T. L., Flannery M. S., Aruffo A., Kaji E. H., Gorn A., Kolakowski L. F., Jr, Lodish H. F., Goldring S. R. Expression cloning of an adenylate cyclase-coupled calcitonin receptor. Science. 1991 Nov 15;254(5034):1022–1024. doi: 10.1126/science.1658940. [DOI] [PubMed] [Google Scholar]
  21. Mathews L. S., Vale W. W. Expression cloning of an activin receptor, a predicted transmembrane serine kinase. Cell. 1991 Jun 14;65(6):973–982. doi: 10.1016/0092-8674(91)90549-e. [DOI] [PubMed] [Google Scholar]
  22. Mayo K. E. Molecular cloning and expression of a pituitary-specific receptor for growth hormone-releasing hormone. Mol Endocrinol. 1992 Oct;6(10):1734–1744. doi: 10.1210/mend.6.10.1333056. [DOI] [PubMed] [Google Scholar]
  23. Nishimura E., Billestrup N., Perrin M., Vale W. Identification and characterization of a pituitary corticotropin-releasing factor binding protein by chemical cross-linking. J Biol Chem. 1987 Sep 25;262(27):12893–12896. [PubMed] [Google Scholar]
  24. Okamoto T., Murayama Y., Hayashi Y., Inagaki M., Ogata E., Nishimoto I. Identification of a Gs activator region of the beta 2-adrenergic receptor that is autoregulated via protein kinase A-dependent phosphorylation. Cell. 1991 Nov 15;67(4):723–730. doi: 10.1016/0092-8674(91)90067-9. [DOI] [PubMed] [Google Scholar]
  25. Orth D. N. Corticotropin-releasing hormone in humans. Endocr Rev. 1992 May;13(2):164–191. doi: 10.1210/edrv-13-2-164. [DOI] [PubMed] [Google Scholar]
  26. Perrin M. H., Bilezikjian L. M., Hoeger C., Donaldson C. J., Rivier J., Haas Y., Vale W. W. Molecular and functional characterization of GnRH receptors cloned from rat pituitary and a mouse pituitary tumor cell line. Biochem Biophys Res Commun. 1993 Mar 31;191(3):1139–1144. doi: 10.1006/bbrc.1993.1335. [DOI] [PubMed] [Google Scholar]
  27. Perrin M. H., Haas Y., Rivier J. E., Vale W. W. Corticotropin-releasing factor binding to the anterior pituitary receptor is modulated by divalent cations and guanyl nucleotides. Endocrinology. 1986 Mar;118(3):1171–1179. doi: 10.1210/endo-118-3-1171. [DOI] [PubMed] [Google Scholar]
  28. Potter E., Behan D. P., Fischer W. H., Linton E. A., Lowry P. J., Vale W. W. Cloning and characterization of the cDNAs for human and rat corticotropin releasing factor-binding proteins. Nature. 1991 Jan 31;349(6308):423–426. doi: 10.1038/349423a0. [DOI] [PubMed] [Google Scholar]
  29. Potter E., Behan D. P., Linton E. A., Lowry P. J., Sawchenko P. E., Vale W. W. The central distribution of a corticotropin-releasing factor (CRF)-binding protein predicts multiple sites and modes of interaction with CRF. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4192–4196. doi: 10.1073/pnas.89.9.4192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Reisine T., Heisler S., Hook V. Y., Axelrod J. Multireceptor-induced release of adrenocorticotropin from anterior pituitary tumor cells. Biochem Biophys Res Commun. 1982 Oct 15;108(3):1251–1257. doi: 10.1016/0006-291x(82)92134-9. [DOI] [PubMed] [Google Scholar]
  31. Reisine T., Hoffman A. Desensitization of corticotropin-releasing factor receptors. Biochem Biophys Res Commun. 1983 Mar 29;111(3):919–925. doi: 10.1016/0006-291x(83)91387-6. [DOI] [PubMed] [Google Scholar]
  32. Rosendale B. E., Jarrett D. B., Robinson A. G. Identification of a corticotropin-releasing factor-binding protein in the plasma membrane of AtT-20 mouse pituitary tumor cells and its regulation by dexamethasone. Endocrinology. 1987 Jun;120(6):2357–2366. doi: 10.1210/endo-120-6-2357. [DOI] [PubMed] [Google Scholar]
  33. Sapolsky R., Rivier C., Yamamoto G., Plotsky P., Vale W. Interleukin-1 stimulates the secretion of hypothalamic corticotropin-releasing factor. Science. 1987 Oct 23;238(4826):522–524. doi: 10.1126/science.2821621. [DOI] [PubMed] [Google Scholar]
  34. Sreedharan S. P., Robichon A., Peterson K. E., Goetzl E. J. Cloning and expression of the human vasoactive intestinal peptide receptor. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4986–4990. doi: 10.1073/pnas.88.11.4986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tilders F. J., Schipper J., Lowry P. J., Vermes I. Effect of hypothalamus lesions on the presence of CRF-immunoreactive nerve terminals in the median eminence and on the pituitary-adrenal response to stress. Regul Pept. 1982 Dec;5(1):77–84. doi: 10.1016/0167-0115(82)90077-5. [DOI] [PubMed] [Google Scholar]
  36. Vale W., Spiess J., Rivier C., Rivier J. Characterization of a 41-residue ovine hypothalamic peptide that stimulates secretion of corticotropin and beta-endorphin. Science. 1981 Sep 18;213(4514):1394–1397. doi: 10.1126/science.6267699. [DOI] [PubMed] [Google Scholar]
  37. Wynn P. C., Aguilera G., Morell J., Catt K. J. Properties and regulation of high-affinity pituitary receptors for corticotropin-releasing factor. Biochem Biophys Res Commun. 1983 Jan 27;110(2):602–608. doi: 10.1016/0006-291x(83)91192-0. [DOI] [PubMed] [Google Scholar]

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