Koch, WJHawes, BEAllen, LFLefkowitz, RJ2013-09-101994-12-200027-8424https://hdl.handle.net/10161/7843Stimulation of Gi-coupled receptors leads to the activation of mitogen-activated protein kinases (MAP kinases). In several cell types, this appears to be dependent on the activation of p21ras (Ras). Which G-protein subunit(s) (G alpha or the G beta gamma complex) primarily is responsible for triggering this signaling pathway, however, is unclear. We have demonstrated previously that the carboxyl terminus of the beta-adrenergic receptor kinase, containing its G beta gamma-binding domain, is a cellular G beta gamma antagonist capable of specifically distinguishing G alpha- and G beta gamma-mediated processes. Using this G beta gamma inhibitor, we studied Ras and MAP kinase activation through endogenous Gi-coupled receptors in Rat-1 fibroblasts and through receptors expressed by transiently transfected COS-7 cells. We report here that both Ras and MAP kinase activation in response to lysophosphatidic acid is markedly attenuated in Rat-1 cells stably transfected with a plasmid encoding this G beta gamma antagonist. Likewise in COS-7 cells transfected with plasmids encoding Gi-coupled receptors (alpha 2-adrenergic and M2 muscarinic), the activation of Ras and MAP kinase was significantly reduced in the presence of the coexpressed G beta gamma antagonist. Ras-MAP kinase activation mediated through a Gq-coupled receptor (alpha 1-adrenergic) or the tyrosine kinase epidermal growth factor receptor was unaltered by this G beta gamma antagonist. These results identify G beta gamma as the primary mediator of Ras activation and subsequent signaling via MAP kinase in response to stimulation of Gi-coupled receptors.AnimalsCell LineCercopithecus aethiopsCyclic AMP-Dependent Protein KinasesEnzyme ActivationGTP-Binding ProteinsGuanosine TriphosphateIn Vitro TechniquesMitogen-Activated Protein Kinase 1Peptide FragmentsProtein-Serine-Threonine KinasesProtein-Tyrosine KinasesProto-Oncogene Proteins p21(ras)RatsReceptors, Adrenergic, betaReceptors, Cell SurfaceReceptors, G-Protein-CoupledReceptors, Lysophosphatidic AcidSignal Transductionbeta-Adrenergic Receptor KinasesJournal article