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Originally published In Press as doi:10.1074/jbc.M200946200 on March 29, 2002

J. Biol. Chem., Vol. 277, Issue 23, 20903-20910, June 7, 2002
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Protein Kinase C Induces Actin Reorganization via a Src- and Rho-dependent Pathway*

Dominique BrandtDagger , Mario Gimona§, Meike HillmannDagger , Hermann HallerDagger , and Harald MischakDagger

From the Dagger  Medizinische Hochschule Hannover, Department of Nephrology, 30625 Hannover, Germany and the § Austrian Academy of Sciences, Institute of Molecular Biology, Department of Cell Biology, A-5020 Salzburg, Austria

We have investigated the mechanism of PKC-induced actin reorganization in A7r5 vascular smooth muscle cells. PKC activation by 12-O-tetradecanoylphorbol-13-acetate induces the disassembly of actin stress fibers concomitant with the appearance of membrane ruffles. PKC also induces rapid tyrosine phosphorylation in these cells. As we could show, utilizing the Src-specific inhibitor PP2 and a kinase-deficient c-Src mutant, actin reorganization is dependent on PKC-induced Src activation. Subsequently, the activity of the small G-protein RhoA is decreased, whereas Rac and Cdc42 activities remain unchanged. Disassembly of actin stress fibers could also be observed using the Rho kinase-specific inhibitor Y-27632, indicating that the decrease in RhoA activity on its own is responsible for actin reorganization. In addition, we show that tyrosine phosphorylation of p190RhoGAP is increased upon 12-O-tetradecanoylphorbol-13-acetate stimulation, directly linking Src activation to a decrease in RhoA activity. Our data provide substantial evidence for a model elucidating the molecular mechanisms of PKC-induced actin rearrangements.

* This work was supported by Grants Mi 489/6-1 and SFB TR-2 from the Deutsche Forschungsgemeinsschaft (to H. M.) and by a grant from the Austrian Science Fund (to M. G.).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.

To whom correspondence should be addressed: Medizinische Hochschule Hannover, Dept. Nephrology, Feodor Lynen Str. 5, 30625 Hannover, Germany. Tel.: 49-511-55474413; Fax: 49-511-55474431; E-mail: mischak@gmx.net.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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