XB-ART-41785
PLoS One
July 23, 2010;
5
(7):
e11489.
The human gonadotropin releasing hormone type I receptor is a functional intracellular GPCR expressed on the nuclear membrane.
Re M
,
Pampillo M
,
Savard M
,
Dubuc C
,
McArdle CA
,
Millar RP
,
Conn PM
,
Gobeil F
,
Bhattacharya M
,
Babwah AV
.
Abstract
The mammalian type I gonadotropin releasing hormone receptor (GnRH-R) is a structurally unique G protein-coupled receptor (GPCR) that lacks cytoplasmic tail sequences and displays inefficient plasma membrane expression (PME). Compared to its murine counterparts, the primate type I receptor is inefficiently folded and retained in the endoplasmic reticulum (ER) leading to a further reduction in PME. The decrease in PME and concomitant increase in intracellular localization of the mammalian GnRH-RI led us to characterize the spatial distribution of the human and mouse GnRH receptors in two human cell lines, HEK 293 and HTR-8/SVneo. In both human cell lines we found the receptors were expressed in the cytoplasm and were associated with the ER and nuclear membrane. A molecular analysis of the receptor protein sequence led us to identify a putative monopartite nuclear localization sequence (NLS) in the first intracellular loop of GnRH-RI. Surprisingly, however, neither the deletion of the NLS nor the addition of the Xenopus GnRH-R cytoplasmic tail sequences to the human receptor altered its spatial distribution. Finally, we demonstrate that GnRH treatment of nuclei isolated from HEK 293 cells expressing exogenous GnRH-RI triggers a significant increase in the acetylation and phosphorylation of histone H3, thereby revealing that the nuclear-localized receptor is functional. Based on our findings, we conclude that the mammalian GnRH-RI is an intracellular GPCR that is expressed on the nuclear membrane. This major and novel discovery causes us to reassess the signaling potential of this physiologically and clinically important receptor.
PubMed ID: 20628612
PMC ID: PMC2900216
Article link: PLoS One
Grant support: [+]
HD-18185 NICHD NIH HHS , HD-19899 NICHD NIH HHS , MOP#89832 Canadian Institutes of Health Research , RR-00163 NCRR NIH HHS , P51 OD011092 NIH HHS , R01 HD019899 NICHD NIH HHS , U54 HD018185 NICHD NIH HHS , P51 RR000163 NCRR NIH HHS , R37 HD019899 NICHD NIH HHS , K01 RR000163 NCRR NIH HHS , P30 HD018185 NICHD NIH HHS
Species referenced: Xenopus laevis
Genes referenced: canx cltc cstb gnrh1 golga2 gprc6a lmna
Article Images: [+] show captions
References [+] :
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Babwah, Protein kinase C isoform-specific differences in the spatial-temporal regulation and decoding of metabotropic glutamate receptor1a-stimulated second messenger responses. 2003, Pubmed
Bhattacharya, Ral and phospholipase D2-dependent pathway for constitutive metabotropic glutamate receptor endocytosis. 2004, Pubmed
Bhattacharya, Nuclear localization of prostaglandin E2 receptors. 1999, Pubmed
Bhattacharya, Localization of functional prostaglandin E2 receptors EP3 and EP4 in the nuclear envelope. 1999, Pubmed
Boivin, Functional endothelin receptors are present on nuclei in cardiac ventricular myocytes. 2003, Pubmed
Brothers, Calnexin regulated gonadotropin-releasing hormone receptor plasma membrane expression. 2006, Pubmed
Brothers, Unexpected effects of epitope and chimeric tags on gonadotropin-releasing hormone receptors: implications for understanding the molecular etiology of hypogonadotropic hypogonadism. 2003, Pubmed
Cao, The composition of the beta-2 adrenergic receptor oligomer affects its membrane trafficking after ligand-induced endocytosis. 2004, Pubmed
Cavanagh, Gonadotropin-releasing hormone-regulated chemokine expression in human placentation. 2009, Pubmed
Chelsky, Sequence requirements for synthetic peptide-mediated translocation to the nucleus. 1989, Pubmed , Xenbase
Chen, A functional angiotensin II receptor-GFP fusion protein: evidence for agonist-dependent nuclear translocation. 2000, Pubmed
Cheng, Regulation of human gonadotropin-releasing hormone receptor gene expression in placental cells. 2000, Pubmed
Chou, Cellular localization of gonadotropin-releasing hormone (GnRH) I and GnRH II in first-trimester human placenta and decidua. 2004, Pubmed
Cornea, Gonadotropin-releasing hormone receptor microaggregation. Rate monitored by fluorescence resonance energy transfer. 2001, Pubmed
Eggena, Nuclear angiotensin receptors induce transcription of renin and angiotensinogen mRNA. 1993, Pubmed
Finch, Plasma membrane expression of GnRH receptors: regulation by antagonists in breast, prostate, and gonadotrope cell lines. 2008, Pubmed , Xenbase
Gardner, AKAP79-mediated targeting of the cyclic AMP-dependent protein kinase to the beta1-adrenergic receptor promotes recycling and functional resensitization of the receptor. 2006, Pubmed
Gobeil, G-protein-coupled receptors signalling at the cell nucleus: an emerging paradigm. 2006, Pubmed
Gobeil, Regulation of eNOS expression in brain endothelial cells by perinuclear EP(3) receptors. 2002, Pubmed
Graham, Establishment and characterization of first trimester human trophoblast cells with extended lifespan. 1993, Pubmed
Grosse, Gonadotropin-releasing hormone receptor initiates multiple signaling pathways by exclusively coupling to G(q/11) proteins. 2000, Pubmed
Hislop, Internalization of gonadotropin-releasing hormone receptors (GnRHRs): does arrestin binding to the C-terminal tail target GnRHRs for dynamin-dependent internalization? 2005, Pubmed , Xenbase
Janovick, Evolved regulation of gonadotropin-releasing hormone receptor cell surface expression. 2004, Pubmed
Janovick, Regulation of G protein-coupled receptor trafficking by inefficient plasma membrane expression: molecular basis of an evolved strategy. 2006, Pubmed
Janovick, Refolding of misfolded mutant GPCR: post-translational pharmacoperone action in vitro. 2007, Pubmed
Janovick, Structure-activity relations of successful pharmacologic chaperones for rescue of naturally occurring and manufactured mutants of the gonadotropin-releasing hormone receptor. 2003, Pubmed
Jong, Functional metabotropic glutamate receptors on nuclei from brain and primary cultured striatal neurons. Role of transporters in delivering ligand. 2005, Pubmed
Jong, Nuclear localization of functional metabotropic glutamate receptor mGlu1 in HEK293 cells and cortical neurons: role in nuclear calcium mobilization and development. 2007, Pubmed
Kaiser, Chromosomal localization of the gonadotropin-releasing hormone receptor gene to human chromosome 4q13.1-q21.1 and mouse chromosome 5. 1994, Pubmed
Kakar, Cloning, sequencing, and expression of human gonadotropin releasing hormone (GnRH) receptor. 1992, Pubmed
Knollman, Multiple G proteins compete for binding with the human gonadotropin releasing hormone receptor. 2008, Pubmed
Leaños-Miranda, Receptor-misrouting: an unexpectedly prevalent and rescuable etiology in gonadotropin-releasing hormone receptor-mediated hypogonadotropic hypogonadism. 2002, Pubmed
Leaños-Miranda, In vitro coexpression and pharmacological rescue of mutant gonadotropin-releasing hormone receptors causing hypogonadotropic hypogonadism in humans expressing compound heterozygous alleles. 2005, Pubmed
Lee, Agonist-independent nuclear localization of the Apelin, angiotensin AT1, and bradykinin B2 receptors. 2004, Pubmed
Lu, Angiotensin II-induced nuclear targeting of the angiotensin type 1 (AT1) receptor in brain neurons. 1998, Pubmed
Mangia, Gonadotropin releasing hormone receptor expression in primary breast cancer: comparison of immunohistochemical, radioligand and Western blot analyses. 2002, Pubmed
Maudsley, Gonadotropin-releasing hormone (GnRH) antagonists promote proapoptotic signaling in peripheral reproductive tumor cells by activating a Galphai-coupling state of the type I GnRH receptor. 2004, Pubmed
Maya-Núñez, Combined modification of intracellular and extracellular loci on human gonadotropin-releasing hormone receptor provides a mechanism for enhanced expression. 2001, Pubmed
Millar, Luteinizing hormone-releasing hormone (LH-RH) binding to purified rat pituitary nuclei. 1983, Pubmed
Morel, Binding and internalization of native gonadoliberin (GnRH) by anterior pituitary gonadotrophs of the rat. A quantitative autoradiographic study after cryoultramicrotomy. 1987, Pubmed
O'Malley, Activation of metabotropic glutamate receptor mGlu5 on nuclear membranes mediates intranuclear Ca2+ changes in heterologous cell types and neurons. 2003, Pubmed
Petrovska, Addition of a signal peptide sequence to the alpha1D-adrenoceptor gene increases the density of receptors, as determined by [3H]-prazosin binding in the membranes. 2005, Pubmed
Petäjä-Repo, Ligands act as pharmacological chaperones and increase the efficiency of delta opioid receptor maturation. 2002, Pubmed
Pietilä, Inefficient maturation of the rat luteinizing hormone receptor. A putative way to regulate receptor numbers at the cell surface. 2005, Pubmed
Saito, RTP family members induce functional expression of mammalian odorant receptors. 2004, Pubmed
Salahpour, Homodimerization of the beta2-adrenergic receptor as a prerequisite for cell surface targeting. 2004, Pubmed
Savard, Expression of endogenous nuclear bradykinin B2 receptors mediating signaling in immediate early gene activation. 2008, Pubmed
Sedgley, Intracellular gonadotropin-releasing hormone receptors in breast cancer and gonadotrope lineage cells. 2006, Pubmed , Xenbase
Spencer, Subcellular localization of prostaglandin endoperoxide H synthases-1 and -2 by immunoelectron microscopy. 1998, Pubmed
Szende, The concentration of LH-RH receptors in the nuclei of pancreatic cancer cells. Effect of (D-Trp6)LH-RH on tumor-bearing Syrian golden hamsters. 1995, Pubmed
Tetsuka, The basic residues in the membrane-proximal C-terminal tail of the rat melanin-concentrating hormone receptor 1 are required for receptor function. 2004, Pubmed
Uberti, Heterodimerization with beta2-adrenergic receptors promotes surface expression and functional activity of alpha1D-adrenergic receptors. 2005, Pubmed
Vadakkadath Meethal, Identification of a gonadotropin-releasing hormone receptor orthologue in Caenorhabditis elegans. 2007, Pubmed
Vandenberghe, Interaction with the unfolded protein response reveals a role for stargazin in biosynthetic AMPA receptor transport. 2005, Pubmed
Venkatesan, A membrane-proximal basic domain and cysteine cluster in the C-terminal tail of CCR5 constitute a bipartite motif critical for cell surface expression. 2001, Pubmed
Wang, NHERF1 regulates parathyroid hormone receptor membrane retention without affecting recycling. 2007, Pubmed
Watson, Nuclear localization of the type 1 parathyroid hormone/parathyroid hormone-related peptide receptor in MC3T3-E1 cells: association with serum-induced cell proliferation. 2000, Pubmed
Watson, Nuclear localization of the type 1 PTH/PTHrP receptor in rat tissues. 2000, Pubmed