The Rab GTPase-Activating Protein AS160 as a Common Regulator of Insulin- and Gαq-Mediated Intracellular GLUT4 Vesicle Distribution

Akt substrate of 160kDa (AS 160) is a Rab GTPase activating protein (GAP) and was recently identified as a component of the insulin signaling pathway of glucose transporter type 4 (GLUT4) translocation. We and others, previously reported that the activation of G alpha q protein-coupled receptors (G alpha qPCRs) also stimulated GLUT4 translocation and glucose uptake in several cell lines. Here, we report that the activation of G alpha qPCRs also promoted phosphorylation of AS 160 by the 5'-AMP activated protein kinase (AMPK). The suppression of AS160 phosphorylation by the siRNA mediated AMPK alpha 1 subunit knockdown promoted GLUT4 vesicle retention in intracellular compartments. This suppression did not affect the ratio of non-induced cell surface GLUT4 to G alpha q-induced it. Rat 3Y1 cells lacking AS 160 did not show insulin-induced GLUT4 translocation. The cells stably expressing GLUT4 revealed GLUT4 vesicles that were mainly localized in the perinuclear region and less frequently on the cell surface. After expression of exogenous AS 160, GLUT4 on the cell surface decreased and GLUT4 vesicles were redistributed throughout the cytoplasm. Although PMA-induced or sodium fluoride-induced GLUT4 translocation was significantly increased in these cells, insulin did not affect GLUT4 translocation. These results suggest that AS 160 is a common regulator of insulin- and G alpha qPCR activation-mediated GLUT4 distribution in the cells.