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J. Biol. Chem., Vol. 276, Issue 52, 49331-49336, December 28, 2001
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From the Department of Physiology and Biophysics, University of
Iowa, Iowa City, Iowa 52242
Incubation of isolated GLUT4-containing vesicles
with Xenopus oocyte extracts resulted in a guanosine
5'-[
Insulin Stimulates Actin Comet Tails on Intracellular
GLUT4-containing Compartments in Differentiated 3T3L1
Adipocytes*,
-thio]triphosphate (GTPS) 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/
VCA. Preparations of
gently sheared (broken) 3T3L1 adipocytes also displayed GTPS 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 Clostridum difficile toxin B, demonstrating a
specific role for a Rho family member small GTP-binding protein.
Expression of N-WASP/VCA 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.
*
This work was supported by Grants DK33823, DK59291, and
DK25925 from the National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
The on-line version of this article (available at
http://www.jbc.org) contains two movies.
To whom correspondence should be addressed. Tel.:
319-335-7823; Fax: 319-335-7886; E-mail:
Jeffrey-Pessin@uiowa.edu.
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