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Salmanian, S, Najafi, SM, Rafipour, M, Arjomand, MR, Shahheydari, H, Ansari, S, Kashkooli, L, Rasouli, SJ, Jazi, MS and Minaei, T (2010) Regulation of GSK-3beta and beta-Catenin by Galphaq in HEK293T cells. Biochem. Biophys. Res. Commun. 395:577-82


Recent studies have shown that heterotrimeric G proteins are involved in the regulation of the canonical Wnt/beta-Catenin pathway. However, the mechanism(s) behind this involvement is (are) poorly understood. Our previous results have shown that activation of Galphaq in Xenopus oocytes leads to inhibition of GSK-3beta and stabilization of the beta-Catenin protein, suggesting that Galphaq might stabilize beta-Catenin via inhibition of GSK-3beta. In this study, we have observed similar results in HEK293T cells. In these cells optimal activation of endogenous Galphaq by expressing M3-muscarinic acetylcholine receptor (with or without carbachol treatment), or exposing the cells to thrombin led to an increase of 2 to 3-fold in endogenous cytoplasmic beta-Catenin protein levels. In addition, expression of the activated mutant of Galphaq (GalphaqQL) dramatically enhanced accumulation of exogenous beta-Catenin with no effect on beta-catenin (CTNNB1) gene transcription. The Galphaq-mediated cellular accumulation of beta-Catenin was blocked by expression of a minigene encoding a Galphaq specific inhibitory peptide but not by a minigene encoding a Galphas blocking peptide. Also, expression of GalphaqQL led to a significant reduction in GSK-3beta kinase activity, supporting the idea that the positive role of Galphaq signaling in inducing cellular accumulation of beta-Catenin is mediated through inhibition of GSK-3beta.


PubMed Online version:10.1016/j.bbrc.2010.04.087


Cell Line; GTP-Binding Protein alpha Subunits, Gq-G11/genetics; GTP-Binding Protein alpha Subunits, Gq-G11/metabolism; Glycogen Synthase Kinase 3/antagonists & inhibitors; Glycogen Synthase Kinase 3/genetics; Glycogen Synthase Kinase 3/metabolism; Humans; Receptor, Muscarinic M3/agonists; Receptor, Muscarinic M3/biosynthesis; Transcription, Genetic; Wnt Proteins/metabolism; beta Catenin/genetics; beta Catenin/metabolism