A critical role of the small GTPase Rac1 in Akt2‐mediated GLUT4 translocation in mouse skeletal muscle

Insulin promotes glucose uptake in skeletal muscle by inducing the translocation of the glucose transporter GLUT4 to the plasma membrane. The serine/threonine kinase Akt2 has been implicated as a key regulator of this insulin action. However, the mechanisms whereby Akt2 regulates multiple steps of GLUT4 translocation remain incompletely understood. Recently, the small GTPase Rac1 has been identified as a skeletal muscle‐specific regulator of insulin‐stimulated glucose uptake. Here, we show that Rac1 is a critical downstream component of the Akt2 pathway in mouse skeletal muscle as well as cultured myocytes. GLUT4 translocation induced by constitutively activated Akt2 was totally dependent on the expression of Rac1 in L6 myocytes. Moreover, we observed the activation of Rac1 when constitutively activated Akt2 was ectopically expressed. Constitutively activated Akt2‐triggered Rac1 activation was diminished by knockdown of FLJ00068, a guanine nucleotide exchange factor for Rac1. Knockdown of Akt2, on the other hand, markedly reduced Rac1 activation by a constitutively activated mutant of phosphoinositide 3–kinase. In mouse skeletal muscle, constitutively activated mutants of Akt2 and phosphoinositide 3‐kinase, when ectopically expressed, induced GLUT4 translocation. Muscle‐specific rac1 knockout markedly diminished Akt2‐ or phosphoinositide 3‐kinase‐induced GLUT4 translocation, highlighting a crucial role of Rac1 downstream of Akt2. Taken together, these results strongly suggest a novel regulatory link between Akt2 and Rac1 in insulin‐dependent signal transduction leading to glucose uptake in skeletal muscle. We show that Rac1 plays a critical role downstream of Akt2 in insulin‐dependent glucose uptake signaling in skeletal muscle. A signaling cascade composed of phosphoinositide 3‐kinase, Akt2, and Rac1 for GLUT4 translocation was demonstrated in cultured myocytes. Furthermore, constitutively activated Akt2 and phosphoinositide 3‐kinase induced GLUT4 translocation to the sarcolemma in wild‐type, but not rac1 knockout, mouse skeletal muscle.