Loading…

Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor

The primary structure of the gonadotropin-releasing hormone (GnRH) receptor was determined by sequencing a functional receptor cDNA isolated by expression cloning from an immortalized murine gonadotroph (alpha T3) cell line. Positive clone pools from a cDNA library were detected by screening express...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1992-10, Vol.267 (30), p.21281-21284
Main Authors: Reinhart, J, Mertz, L.M., Catt, K.J.
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533
cites cdi_FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533
container_end_page 21284
container_issue 30
container_start_page 21281
container_title The Journal of biological chemistry
container_volume 267
creator Reinhart, J
Mertz, L.M.
Catt, K.J.
description The primary structure of the gonadotropin-releasing hormone (GnRH) receptor was determined by sequencing a functional receptor cDNA isolated by expression cloning from an immortalized murine gonadotroph (alpha T3) cell line. Positive clone pools from a cDNA library were detected by screening expressed RNA in aequorin-injected Xenopus laevis oocytes, in which receptor-mediated calcium responses were monitored as light emission during stimulation by GnRH. The isolated receptor cDNA encodes a 327-amino acid protein that has seven putative transmembrane regions and is unique among G protein-coupled receptors in that the predicted sequence lacks a carboxyl-terminal cytoplasmic domain. COS-7 cells transfected with the receptor cDNA expressed high affinity binding sites for GnRH and its agonist and antagonist analogs and exhibited calcium responses to GnRH stimulation. These, and the prominent calcium responses of Xenopus oocytes injected with receptor RNA, were inhibited by GnRH antagonists. Northern blot analysis revealed two mRNAs (1.6 and 3.5 kilobases) in alpha T3 cells and in the mouse pituitary gland, and both transcripts were shown to encode functional GnRH receptors when expressed in Xenopus oocytes. In contrast, a single 4.6-kilobase receptor mRNA was present in rat anterior pituitary gland, ovary, and Leydig cells. The absence of a carboxyl-terminal cytoplasmic domain indicates the importance of other regions of the GnRH receptor in agonist-induced signal transduction, and possibly in receptor desensitization and sequestration.
doi_str_mv 10.1016/S0021-9258(19)36602-5
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_73233838</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925819366025</els_id><sourcerecordid>73233838</sourcerecordid><originalsourceid>FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533</originalsourceid><addsrcrecordid>eNqFkUtP3TAUhK2qFb2F_gSkLKqqXYT6EefaqwrxKhKUBSCxsxz7GFwldmoniP57HO4VXWIvvJjvzLFmENon-IBg0v64xpiSWlIuvhH5nbUtpjV_h1YEC1YzTu7eo9Ur8hF9yvkPLqeRZAftEEYFpWKF7GXswcy9TpXpY_DhvtLBVvA0JsjZx1BFV5nj34cVBBPtok8PUA1z8gGq-xi0jVOKow91gh50XoiHmIZY5AQGximmPfTB6T7D5-27i25PT26OftUXV2fnR4cXteGMTTUVkq8NE9R1ayY6IZ3WjEpHJKXWYgmt68A22jLegeTOsgYM11jq1jpdLHbR143vmOLfGfKkBp8N9L0OEOes1owyJsp9CyQtJ01LFke-AU2KOSdwakx-0OmfIlgtNaiXGtSSsSJSvdSgeJnb3y6YuwHs_6lN7kX_stV1Nrp3SQfj8yvWNJgLTgr2c4NBSe3RQ1LZ-FIEWF-ynZSN_o2PPANDyaWt</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16514613</pqid></control><display><type>article</type><title>Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor</title><source>Elsevier ScienceDirect Journals</source><creator>Reinhart, J ; Mertz, L.M. ; Catt, K.J.</creator><creatorcontrib>Reinhart, J ; Mertz, L.M. ; Catt, K.J.</creatorcontrib><description>The primary structure of the gonadotropin-releasing hormone (GnRH) receptor was determined by sequencing a functional receptor cDNA isolated by expression cloning from an immortalized murine gonadotroph (alpha T3) cell line. Positive clone pools from a cDNA library were detected by screening expressed RNA in aequorin-injected Xenopus laevis oocytes, in which receptor-mediated calcium responses were monitored as light emission during stimulation by GnRH. The isolated receptor cDNA encodes a 327-amino acid protein that has seven putative transmembrane regions and is unique among G protein-coupled receptors in that the predicted sequence lacks a carboxyl-terminal cytoplasmic domain. COS-7 cells transfected with the receptor cDNA expressed high affinity binding sites for GnRH and its agonist and antagonist analogs and exhibited calcium responses to GnRH stimulation. These, and the prominent calcium responses of Xenopus oocytes injected with receptor RNA, were inhibited by GnRH antagonists. Northern blot analysis revealed two mRNAs (1.6 and 3.5 kilobases) in alpha T3 cells and in the mouse pituitary gland, and both transcripts were shown to encode functional GnRH receptors when expressed in Xenopus oocytes. In contrast, a single 4.6-kilobase receptor mRNA was present in rat anterior pituitary gland, ovary, and Leydig cells. The absence of a carboxyl-terminal cytoplasmic domain indicates the importance of other regions of the GnRH receptor in agonist-induced signal transduction, and possibly in receptor desensitization and sequestration.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(19)36602-5</identifier><identifier>PMID: 1328228</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Base Sequence ; Biological and medical sciences ; Blotting, Northern ; calcium ; cDNA ; Cell Line ; Cell receptors ; Cell structures and functions ; cloning ; Cloning, Molecular ; DNA ; expression ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; genes ; gonadotropin-releasing hormone ; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors ; Mice ; Molecular and cellular biology ; Molecular Sequence Data ; nucleotide sequence ; Plasmids ; predictions ; receptors ; Receptors, LHRH - genetics ; response ; RNA, Messenger - genetics ; Xenopus laevis</subject><ispartof>The Journal of biological chemistry, 1992-10, Vol.267 (30), p.21281-21284</ispartof><rights>1992 © 1992 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533</citedby><cites>FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0021925819366025$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3549,27924,27925,45780</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=4405851$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1328228$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Reinhart, J</creatorcontrib><creatorcontrib>Mertz, L.M.</creatorcontrib><creatorcontrib>Catt, K.J.</creatorcontrib><title>Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The primary structure of the gonadotropin-releasing hormone (GnRH) receptor was determined by sequencing a functional receptor cDNA isolated by expression cloning from an immortalized murine gonadotroph (alpha T3) cell line. Positive clone pools from a cDNA library were detected by screening expressed RNA in aequorin-injected Xenopus laevis oocytes, in which receptor-mediated calcium responses were monitored as light emission during stimulation by GnRH. The isolated receptor cDNA encodes a 327-amino acid protein that has seven putative transmembrane regions and is unique among G protein-coupled receptors in that the predicted sequence lacks a carboxyl-terminal cytoplasmic domain. COS-7 cells transfected with the receptor cDNA expressed high affinity binding sites for GnRH and its agonist and antagonist analogs and exhibited calcium responses to GnRH stimulation. These, and the prominent calcium responses of Xenopus oocytes injected with receptor RNA, were inhibited by GnRH antagonists. Northern blot analysis revealed two mRNAs (1.6 and 3.5 kilobases) in alpha T3 cells and in the mouse pituitary gland, and both transcripts were shown to encode functional GnRH receptors when expressed in Xenopus oocytes. In contrast, a single 4.6-kilobase receptor mRNA was present in rat anterior pituitary gland, ovary, and Leydig cells. The absence of a carboxyl-terminal cytoplasmic domain indicates the importance of other regions of the GnRH receptor in agonist-induced signal transduction, and possibly in receptor desensitization and sequestration.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Northern</subject><subject>calcium</subject><subject>cDNA</subject><subject>Cell Line</subject><subject>Cell receptors</subject><subject>Cell structures and functions</subject><subject>cloning</subject><subject>Cloning, Molecular</subject><subject>DNA</subject><subject>expression</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>genes</subject><subject>gonadotropin-releasing hormone</subject><subject>Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><subject>Molecular Sequence Data</subject><subject>nucleotide sequence</subject><subject>Plasmids</subject><subject>predictions</subject><subject>receptors</subject><subject>Receptors, LHRH - genetics</subject><subject>response</subject><subject>RNA, Messenger - genetics</subject><subject>Xenopus laevis</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><recordid>eNqFkUtP3TAUhK2qFb2F_gSkLKqqXYT6EefaqwrxKhKUBSCxsxz7GFwldmoniP57HO4VXWIvvJjvzLFmENon-IBg0v64xpiSWlIuvhH5nbUtpjV_h1YEC1YzTu7eo9Ur8hF9yvkPLqeRZAftEEYFpWKF7GXswcy9TpXpY_DhvtLBVvA0JsjZx1BFV5nj34cVBBPtok8PUA1z8gGq-xi0jVOKow91gh50XoiHmIZY5AQGximmPfTB6T7D5-27i25PT26OftUXV2fnR4cXteGMTTUVkq8NE9R1ayY6IZ3WjEpHJKXWYgmt68A22jLegeTOsgYM11jq1jpdLHbR143vmOLfGfKkBp8N9L0OEOes1owyJsp9CyQtJ01LFke-AU2KOSdwakx-0OmfIlgtNaiXGtSSsSJSvdSgeJnb3y6YuwHs_6lN7kX_stV1Nrp3SQfj8yvWNJgLTgr2c4NBSe3RQ1LZ-FIEWF-ynZSN_o2PPANDyaWt</recordid><startdate>19921025</startdate><enddate>19921025</enddate><creator>Reinhart, J</creator><creator>Mertz, L.M.</creator><creator>Catt, K.J.</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7Z</scope><scope>M81</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19921025</creationdate><title>Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor</title><author>Reinhart, J ; Mertz, L.M. ; Catt, K.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Northern</topic><topic>calcium</topic><topic>cDNA</topic><topic>Cell Line</topic><topic>Cell receptors</topic><topic>Cell structures and functions</topic><topic>cloning</topic><topic>Cloning, Molecular</topic><topic>DNA</topic><topic>expression</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>genes</topic><topic>gonadotropin-releasing hormone</topic><topic>Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors</topic><topic>Mice</topic><topic>Molecular and cellular biology</topic><topic>Molecular Sequence Data</topic><topic>nucleotide sequence</topic><topic>Plasmids</topic><topic>predictions</topic><topic>receptors</topic><topic>Receptors, LHRH - genetics</topic><topic>response</topic><topic>RNA, Messenger - genetics</topic><topic>Xenopus laevis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Reinhart, J</creatorcontrib><creatorcontrib>Mertz, L.M.</creatorcontrib><creatorcontrib>Catt, K.J.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 1</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Reinhart, J</au><au>Mertz, L.M.</au><au>Catt, K.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1992-10-25</date><risdate>1992</risdate><volume>267</volume><issue>30</issue><spage>21281</spage><epage>21284</epage><pages>21281-21284</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>The primary structure of the gonadotropin-releasing hormone (GnRH) receptor was determined by sequencing a functional receptor cDNA isolated by expression cloning from an immortalized murine gonadotroph (alpha T3) cell line. Positive clone pools from a cDNA library were detected by screening expressed RNA in aequorin-injected Xenopus laevis oocytes, in which receptor-mediated calcium responses were monitored as light emission during stimulation by GnRH. The isolated receptor cDNA encodes a 327-amino acid protein that has seven putative transmembrane regions and is unique among G protein-coupled receptors in that the predicted sequence lacks a carboxyl-terminal cytoplasmic domain. COS-7 cells transfected with the receptor cDNA expressed high affinity binding sites for GnRH and its agonist and antagonist analogs and exhibited calcium responses to GnRH stimulation. These, and the prominent calcium responses of Xenopus oocytes injected with receptor RNA, were inhibited by GnRH antagonists. Northern blot analysis revealed two mRNAs (1.6 and 3.5 kilobases) in alpha T3 cells and in the mouse pituitary gland, and both transcripts were shown to encode functional GnRH receptors when expressed in Xenopus oocytes. In contrast, a single 4.6-kilobase receptor mRNA was present in rat anterior pituitary gland, ovary, and Leydig cells. The absence of a carboxyl-terminal cytoplasmic domain indicates the importance of other regions of the GnRH receptor in agonist-induced signal transduction, and possibly in receptor desensitization and sequestration.</abstract><cop>Bethesda, MD</cop><pub>Elsevier Inc</pub><pmid>1328228</pmid><doi>10.1016/S0021-9258(19)36602-5</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0021-9258
ispartof The Journal of biological chemistry, 1992-10, Vol.267 (30), p.21281-21284
issn 0021-9258
1083-351X
language eng
recordid cdi_proquest_miscellaneous_73233838
source Elsevier ScienceDirect Journals
subjects Amino Acid Sequence
Animals
Base Sequence
Biological and medical sciences
Blotting, Northern
calcium
cDNA
Cell Line
Cell receptors
Cell structures and functions
cloning
Cloning, Molecular
DNA
expression
Fundamental and applied biological sciences. Psychology
Gene Expression
genes
gonadotropin-releasing hormone
Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors
Mice
Molecular and cellular biology
Molecular Sequence Data
nucleotide sequence
Plasmids
predictions
receptors
Receptors, LHRH - genetics
response
RNA, Messenger - genetics
Xenopus laevis
title Molecular cloning and expression of cDNA encoding the murine gonadotropin-releasing hormone receptor
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T20%3A33%3A51IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Molecular%20cloning%20and%20expression%20of%20cDNA%20encoding%20the%20murine%20gonadotropin-releasing%20hormone%20receptor&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Reinhart,%20J&rft.date=1992-10-25&rft.volume=267&rft.issue=30&rft.spage=21281&rft.epage=21284&rft.pages=21281-21284&rft.issn=0021-9258&rft.eissn=1083-351X&rft.coden=JBCHA3&rft_id=info:doi/10.1016/S0021-9258(19)36602-5&rft_dat=%3Cproquest_cross%3E73233838%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c533t-28957c382fb738b89faa329f1922dd09e6fbed4ad35be95fd34ec5a09a6dfa533%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=16514613&rft_id=info:pmid/1328228&rfr_iscdi=true