title: Activated G alpha q inhibits p110 alpha phosphatidylinositol 3-kinase and Akt.
abstract: Some Gq-coupled receptors have been shown to antagonize growth factor activation of phosphatidylinositol 3-kinase (PI3K) and its downstream effector, Akt. We used a constitutively active Galphaq(Q209L) mutant to explore the effects of Galphaq activation on signaling through the PI3K/Akt pathway. Transient expression of Galphaq(Q209L) in Rat-1 fibroblasts inhibited Akt activation induced by platelet-derived growth factor or insulin treatment. Expression of Galphaq(Q209L) also attenuated Akt activation promoted by coexpression of constitutively active PI3K in human embryonic kidney 293 cells. Galphaq(Q209L) had no effect on the activity of an Akt mutant in which the two regulatory phosphorylation sites were changed to acidic amino acids. Inducible expression of Galphaq(Q209L) in a stably transfected 293 cell line caused a decrease in PI3K activity in p110alpha (but not p110beta) immunoprecipitates. Receptor activation of Galphaq also selectively inhibited PI3K activity in p110alpha immunoprecipitates. Active Galphaq still inhibited PI3K/Akt in cells pretreated with the phospholipase C inhibitor U73122. Finally, Galphaq(Q209L) co-immunoprecipitated with the p110alpha-p85alpha PI3K heterodimer from lysates of COS-7 cells expressing these proteins, and incubation of immunoprecipitated Galphaq(Q209L) with purified recombinant p110alpha-p85alpha in vitro led to a decrease in PI3K activity. These results suggest that agonist binding to Gq-coupled receptors blocks Akt activation via the release of active Galphaq subunits that inhibit PI3K. The inhibitory mechanism seems to be independent of phospholipase C activation and might involve an inhibitory interaction between Galphaq and p110alpha PI3K.
authors.affiliation: Research Service, Department of Veterans Affairs Medical Center, Northport, New York 11768, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Ballou
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Lin
authors.forename: Gaofeng
authors.initials: G
authors.lastname: Fan
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Jiang
authors.forename: Richard Z
authors.initials: RZ
authors.lastname: Lin
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D006801:Humans; D051379:Mice; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D021122:Protein Subunits / Q000037:antagonists & inhibitors; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors*; D051057:Proto-Oncogene Proteins c-akt; D011972:Receptor, Insulin / Q000502:physiology; D018340:Receptors, Adrenergic, alpha-1 / Q000502:physiology; D014162:Transfection; D010738:Type C Phospholipases
chemical_list: C547713:ADRA1A protein, human; C547714:Adra1a protein, mouse; C547715:Adra1a protein, rat; D000081082:Phosphoinositide-3 Kinase Inhibitors; D021122:Protein Subunits; D011518:Proto-Oncogene Proteins; D018340:Receptors, Adrenergic, alpha-1; D011972:Receptor, Insulin; C494918:AKT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D010738:Type C Phospholipases; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins