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RGS Protein Specificity Towards Gq- and Gi/o-Mediated ERK 1/2 and Akt Activation, in vitro

  • Anger, Thomas (Department for Cardiology, Friedrich-Alexander University Erlangen) ;
  • Klintworth, Nils (Department for Cardiology, Friedrich-Alexander University Erlangen) ;
  • Stumpf, Christian (Department for Cardiology, Friedrich-Alexander University Erlangen) ;
  • Daniel, Werner G. (Department for Cardiology, Friedrich-Alexander University Erlangen) ;
  • Mende, Ulrike (Cardiovascular Research Center at Brown Medical School) ;
  • Garlichs, Christoph D. (Department for Cardiology, Friedrich-Alexander University Erlangen)
  • Published : 2007.11.30
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Abstract

Extracellular Regulated Kinases (ERK) and Protein Kinase B (Akt) are intermediaries in relaying extracellular growth signals to intracellular targets. Each pathway can become activated upon stimulation of G protein-coupled receptors mediated by $G_q$ and $G_{i/o}$ proteins subjected to regulation by RGS proteins. The goal of the study was to delineate the specificity in which cardiac RGS proteins modulate $G_{q^-}$ and $G_{i/o}$-induced ERK and Akt phosphorylation. To isolate $G_{q^-}$ and $G_{i/o}$-mediated effects, we exclusively expressed muscarinic $M_2$ or $M_3$ receptors in COS-7 cells. Western blot analyses demonstrated increase of phosphorylation of ERK 1.7-/3.3-fold and Akt 2.4-/6-fold in $M_{2^-}/M_{3^-}$ expressing cells through carbachol stimulation. In co-expressions, $M_3/G_q$-induced activation of Akt was exclusively blunted through RGS3s/RGS3, whereas activation of ERK was inhibited additionally through RGS2/RGS5. $M_2/G_{i/o}$ induced Akt activation was inhibited by all RGS proteins tested. RGS2 had no effect on $M_2/G_{i/o}$-induced ERK activation. The high degree of specificity in RGS proteins-depending modulation of $G_{q^-}$ and $G_{i/o}$-mediated ERK and Akt activation in the muscarinic network cannot merely be attributed exclusively to RGS protein selectivity towards $G_q$ or $G_{i/o}$ proteins. Counter-regulatory mechanisms and inter-signaling cross-talk may alter the sensitivity of GPCR-induced ERK and Akt activation to RGS protein regulation.

Keywords

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