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Papers In Press, published online ahead of print October 17, 2001
Department of Physiology & Biophysics, The University of Iowa, Iowa City, IA 52242-1109
Corresponding Author: jeffrey-pessin{at}uiowa.edu
Incubation of isolated GLUT4-containing vesicles with Xenopus oocyte extracts resulted in a GTPgS and sodium orthovanadate stimulation of actin comet tails. The in vitro actin-based GLUT4 vesicle motility was inhibited by both latrunculin B and a dominant-interfering N-WASP mutant, N-WASP/DVCA. Preparations of gently sheared (broken) 3T3L1 adipocytes also displayed GTPgS and sodium orthovanadate stimulation of actin comet tails on GLUT4 intracellular compartments. Furthermore, insulin pretreatment of intact adipocytes prior to gently shearing also resulted in a marked increase in actin polymerization and actin comet tailing on GLUT4 vesicles. In addition, the insulin stimulation of actin comet tails was completely inhibited by C. difficile toxin B, demonstrating a specific role for a Rho family member small GTP binding protein. Expression of N-WASP/DVCA in intact cells had little effect on adipocyte cortical actin but partially inhibited insulin-stimulated GLUT4 translocation. Taken together, these data demonstrate that insulin can induce GLUT4 vesicle actin comet tails that are necessary for the efficient translocation of GLUT4 from intracellular storage sites to the plasma membrane.
J. Biol. Chem, 10.1074/jbc.M109657200
Submitted on October 5, 2001
Revised on October 17, 2001
Accepted on October 16, 2001
Insulin stimulates actin comet tails on intracellular GLUT4-containing compartments in differentiated 3T3L1 adipocytes
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