Expression Cloning of a Human Corticotropin-Releasing-Factor Receptor
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...
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| Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 19; pp. 8967 - 8971 |
|---|---|
| Main Authors | , , , |
| 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 | |
| Online Access | Get full text |
| ISSN | 0027-8424 1091-6490 |
| DOI | 10.1073/pnas.90.19.8967 |
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| 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 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. |
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| AbstractList | 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 cloning of a cDNA coding for a corticotropin-releasing factor (CRF) receptor from a human corticotropic tumor library is reported. A variant of the receptor was produced by alternative splicing. 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. 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. 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.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. |
| Author | Ruoping Chen Wylie W. Vale Marilyn H. Perrin Kathy A. Lewis |
| AuthorAffiliation | Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 |
| AuthorAffiliation_xml | – name: Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 |
| Author_xml | – sequence: 1 givenname: R surname: Chen fullname: Chen, R organization: Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 – sequence: 2 givenname: K A surname: Lewis fullname: Lewis, K A organization: Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 – sequence: 3 givenname: M H surname: Perrin fullname: Perrin, M H organization: Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 – sequence: 4 givenname: W W surname: Vale fullname: Vale, W W organization: Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, CA 92037 |
| BackLink | http://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3786744$$DView record in Pascal Francis https://www.ncbi.nlm.nih.gov/pubmed/7692441$$D View this record in MEDLINE/PubMed |
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| CODEN | PNASA6 |
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| Copyright | Copyright 1993 The National Academy of Sciences of the United States of America 1994 INIST-CNRS Copyright National Academy of Sciences Oct 1, 1993 |
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| Issue | 19 |
| Keywords | 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 |
| Language | English |
| License | CC BY 4.0 |
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| PublicationTitle | Proceedings of the National Academy of Sciences - PNAS |
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| Snippet | Corticotropin-releasing factor (CRF) is the principal neuroregulator of the hypothalamic-pituitary-adrenocortical axis and plays an important role in... The cloning of a cDNA coding for a corticotropin-releasing factor (CRF) receptor from a human corticotropic tumor library is reported. A variant of the... |
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| SubjectTerms | 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 |
| Title | Expression Cloning of a Human Corticotropin-Releasing-Factor Receptor |
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