Gα(h) (or transglutaminase-2 (TG2)) is an atypical guanine nucleotide binding-protein that associates with G protein-coupled receptors. TG2 also exerts transglutaminase activity that catalyzes posttranslational protein cross-linking with the formation of ε-(γ-glutamyl) lysine or (γ-glutamyl) polyamine bonds. Here, the role of Gα(h)/TG2 in signal transduction in glial cells was examined in detail. In 1321N1 human astrocytoma cells that lack Gα(h)/TG2, overexpression of Gα(h)/TG2 caused an enhancement of cAMP accumulation stimulated with the β-adrenergic receptor agonist, isoproterenol, or the adenylylcyclase activator, forskolin. This cAMP-enhancement was reversed by the TG2 inhibitor, ERW1069. In rat C6 glioma cells that express endogenous Gα(h)/TG2, cAMP accumulation induced by isoproterenol or forskolin was significantly inhibited by overexpression of Gα(h)/TG2-C277V, a dominant-negative mutant that lacks transglutaminase activity, but was not inhibited by the Gα(h)/TG2-S171E mutant that cannot bind GTP/GDP. These results suggest Gα(h)/TG2 potentiates adenylylcyclase activity by its transglutaminase activity and not by its G-protein activity. Gα(h)/TG2 also increased the activities of the cAMP response element and interleukin-6 promoter, accompanied by an of cAMP in both glioma cells. Since adenylylcyclase 8 plays a major role in cAMP production, we focused on post-translational modification of adenylylcyclase 8 by Gα(h)/TG2. Adenylylcyclase 8 is expressed in both 1321N1 and C6 cells; however, Gα(h)/TG2 affected neither adenylylcyclase 8 expression levels, glycosylation, nor dimerization status. In contrast, pentylamine, a substrate of Gα(h)/TG2, was incorporated into adenylylcyclase 8 in a transglutaminase activity-dependent manner. Taking these results together, Gα(h)/TG2 promotes cAMP production accompanied by a modification of adenylylcyclase 8 in glioma cells.
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