Lack of receptor-selective effects of either RGS2, RGS3 or RGS4 on muscarinic M3- and gonadotropin-releasing hormone receptor-mediated signalling through G alpha q/11.
Termination of signalling by G-protein-coupled receptors requires inactivation of the G alpha-subunits of heterotrimeric G-proteins and the re-association of G alpha- and G betagamma-subunits. Inactivation of G alpha-subunits is achieved by the hydrolysis of bound GTP by an intrinsic GTPase activity, which is considerably enhanced by GTPase activating proteins. Regulators of G-protein signalling (RGS) proteins are a large family of GTPase activating proteins, many of which have structures indicating roles beyond GTPase activating protein activity and suggesting that the identity of the RGS protein recruited may also be critical to other aspects of signalling. There is some evidence of selective effects of RGS proteins against different G-protein-coupled receptors coupling to the same signalling pathways and growing evidence of physical interactions between RGS proteins and G-protein-coupled receptors. However, it is unclear as to how common such interactions are and the circumstances under which they are functionally relevant. Here we have examined potential selectivity of RGS2, 3 and 4 against signalling mediated by G alpha q/11-coupled muscarinic M3 receptors and gonadotropin-releasing hormone in an immortalised mouse pituitary cell line. Despite major structural differences between these two receptor types and agonist-dependent phosphorylation of the muscarinic M3- but not gonadotropin-releasing hormone receptor, signalling by both receptors was similarly inhibited by expression of either RGS2 or RGS3, whereas RGS4 has little effect. Thus, at least in these circumstances, RGS protein-dependent inhibition of signalling is not influenced by the nature of the G-protein-coupled receptor through which the signalling is mediated.
Karakoula
Aikaterini
A
Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, United Kingdom.
Tovey
Stephen C
SC
Brighton
Paul J
PJ
Willars
Gary B
GB
D000818:Animals; D015374:Biosensing Techniques; D020013:Calcium Signaling / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D003627:Data Interpretation, Statistical; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000494:pharmacology*; D020690:GTPase-Activating Proteins / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000187:drug effects; D016210:Methacholine Chloride / Q000494:pharmacology; D018721:Muscarinic Agonists / Q000494:pharmacology; D010766:Phosphorylation; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D020710:RGS Proteins / Q000494:pharmacology*; D051381:Rats; D043587:Receptor, Muscarinic M3 / Q000187:drug effects*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D011966:Receptors, LHRH / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects*
D020690:GTPase-Activating Proteins; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018721:Muscarinic Agonists; D016271:Proto-Oncogene Proteins c-myc; D020710:RGS Proteins; C087182:RGS2 protein, human; C485898:RGS3 protein, human; D043587:Receptor, Muscarinic M3; D043562:Receptors, G-Protein-Coupled; D011966:Receptors, LHRH; D016210:Methacholine Chloride; C098799:RGS4 protein; D004247:DNA; D019204:GTP-Binding Proteins; D043802:GTP-Binding Protein alpha Subunits, Gq-G11