Independent of GPCRs and Nucleotide Exchange

Abstract

Sato et al . used a strain of yeast lacking the pheromone receptor to functionally screen mammalian cDNA libraries and identify nonreceptor proteins that activate heterotrimeric guanine nucleotide-binding protein (G protein) signaling. They identified a previously uncharacterized protein that they called activator of G protein signaling (AGS) 8, which was derived from cardiac tissue in a rat model of transient myocardial ischemia. Expression of AGS8 mRNA was increased in ischemic ventricles but not in other models of cardiac dysfunction. The effects of hypoxia were specific: Whereas expression of AGS8 mRNA increased in response to hypoxia in cultured ventricular myocytes, it was unaffected in aortic smooth muscle cells, aortic endothelial cells, and cardiac fibroblasts. AGS8 was functionally active in yeast strains that expressed various Gα subunits, including a mutant Gα i2 believed to remain bound to GDP (rather than exchanging GTP for GDP), but required the presence of Gβγ and downstream components of the mitogen-activated protein kinase signaling pathway (the kinase Ste20 and the scaffolding protein Ste5). Moreover, an in vitro binding assay indicated that AGS8 interacted with Gβγ but not with Gα i1/2 , Gα i3 , Gα o , or Gα s . In COS-7 cells, coexpression of AGS8 with Gβγ and the Gβγ effector PLC-β2 did not inhibit Gβγ-stimulated inositol phosphate production; rather, it reversed inhibition of Gβγ-stimulated inositol phosphate production by Gα 13 . A similar reversal by AGS8 of inhibitory effects of Gα on Gβγ function was apparent in yeast. Thus, AGS8 appears to be a regulator of G protein signaling that is induced in ischemic heart to act through a mechanism involving interaction with Gβγ. M. Sato, M. J. Cismowski, E. Toyota, A. V. Smrcka, P. A. Lucchesi, W. M. Chilian, S. M. Lanier, Identification of a receptor-independent activator of G protein signaling (AGS8) in ischemic heart and its interaction with Gβγ. Proc. Natl. Acad. Sci. U.S.A. 103 , 797-802 (2006). [Abstract] [Full Text]