Expression Cloning of a Human Corticotropin-Releasing-Factor Receptor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 19; pp. 8967 - 8971
• Main Authors: Chen, R, Lewis, K A, Perrin, M H, Vale, W W
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.10.1993; National Acad Sciences; National Academy of Sciences
• Subjects: 1-Methyl-3-isobutylxanthine - pharmacology; Adenoma - metabolism; adrenocorticotropic hormone-releasing hormone receptors; Amino Acid Sequence; Amino acids; Animals; Binding, Competitive; Biochemistry; Biological and medical sciences; Blotting, Northern; Brain - metabolism; cDNA; Cell Line; Cell lines; Cell receptors; Cell structures and functions; Cloning; Cloning, Molecular; Complementary DNA; Cyclic AMP - metabolism; Deoxyribonucleic acid; DNA; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene Library; genes; guanine nucleotide-binding protein; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors; Human cloning; Humans; hypothalamus; Kinetics; Libraries; man; Molecular and cellular biology; Molecular Sequence Data; Myocardium - metabolism; nucleotide sequence; Phosphorylation; Pituitary Gland - metabolism; Poly A - biosynthesis; Poly A - metabolism; predictions; Proteins; Radioligand Assay; Receptors; Receptors, Corticotropin-Releasing Hormone - biosynthesis; Receptors, Corticotropin-Releasing Hormone - genetics; Receptors, Corticotropin-Releasing Hormone - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - metabolism; RNA - biosynthesis; RNA - metabolism; RNA, Messenger - biosynthesis; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; Transfection; Tumor Cells, Cultured; Tumors; Human; Alternative splicing; Ligand binding; Molecular form; Peptide hormone; ACTH-RH; Adenoma; Gene expression; Complementary DNA; Pituitary gland; Hormone releasing factor; Molecular cloning; Hormonal receptor
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.19.8967

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 α-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.