Document Detail


Regulation of GPR54 signaling by GRK2 and {beta}-arrestin.
MedLine Citation:
PMID:  19846537     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Kisspeptin and its receptor, GPR54, are major regulators of the hypothalamic-pituitary-gonadal axis as well as regulators of human placentation and tumor metastases. GPR54 is a G(q/11)-coupled G protein-coupled receptor (GPCR), and activation by kisspeptin stimulates phosphatidy linositol 4, 5-biphosphate hydrolysis, Ca(2+) mobilization, arachidonic acid release, and ERK1/2 MAPK phosphorylation. Physiological evidence suggests that GPR54 undergoes agonist-dependent desensitization, but underlying molecular mechanisms are unknown. Furthermore, very little has been reported on the early events that regulate GPR54 signaling. The lack of information in these important areas led to this study. Here we report for the first time on the role of GPCR serine/threonine kinase (GRK)2 and beta-arrestin in regulating GPR54 signaling in human embryonic kidney (HEK) 293 cells, a model cell system for studying the molecular regulation of GPCRs, and genetically modified MDA MB-231 cells, an invasive breast cancer cell line expressing about 75% less beta-arrestin-2 than the control cell line. Our study reveals that in HEK 293 cells, GPR54 is expressed both at the plasma membrane and intracellularly and also that plasma membrane expression is regulated by cytoplasmic tail sequences. We also demonstrate that GPR54 exhibits constitutive activity, internalization, and association with GRK2 and beta- arrestins-1 and 2 through sequences in the second intracellular loop and cytoplasmic tail of the receptor. We also show that GRK2 stimulates the desensitization of GPR54 in HEK 293 cells and that beta-arrestin-2 mediates GPR54 activation of ERK1/2 in MDA-MB-231 cells. The significance of these findings in developing molecular-based therapies for treating certain endocrine-related disorders is discussed.
Authors:
Macarena Pampillo; Natasha Camuso; Jay E Taylor; Jacob M Szereszewski; Maryse R Ahow; Mateusz Zajac; Robert P Millar; Moshmi Bhattacharya; Andy V Babwah
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-10-21
Journal Detail:
Title:  Molecular endocrinology (Baltimore, Md.)     Volume:  23     ISSN:  1944-9917     ISO Abbreviation:  Mol. Endocrinol.     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-25     Completed Date:  2010-02-22     Revised Date:  2012-06-07    
Medline Journal Info:
Nlm Unique ID:  8801431     Medline TA:  Mol Endocrinol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2060-74     Citation Subset:  IM    
Affiliation:
Children's Health Research Institute, Department of Obstetrics and Gynaecology, The University of Western Ontario, London, Canada.
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MeSH Terms
Descriptor/Qualifier:
Arrestins / metabolism*
Cell Line
Cell Line, Tumor
Cell Membrane / metabolism*
G-Protein-Coupled Receptor Kinase 2 / genetics,  metabolism*
Humans
Immunoprecipitation
Kisspeptins
Microscopy, Confocal
Oligopeptides / pharmacology
Phosphorylation / drug effects,  genetics
Protein Binding
Protein Transport / drug effects
Receptors, G-Protein-Coupled / genetics,  metabolism*
Signal Transduction* / drug effects,  genetics
Tumor Suppressor Proteins / genetics
Grant Support
ID/Acronym/Agency:
MOP 81383//Canadian Institutes of Health Research
Chemical
Reg. No./Substance:
0/Arrestins; 0/KISS1 protein, human; 0/KISS1R protein, human; 0/Kisspeptins; 0/Oligopeptides; 0/Receptors, G-Protein-Coupled; 0/Tumor Suppressor Proteins; 0/beta-arrestin; EC 2.7.11.15/ADRBK1 protein, human; EC 2.7.11.15/G-Protein-Coupled Receptor Kinase 2

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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