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Vol. 10, Issue 8, 2481-2491, August 1999
and
*Department of Molecular Biology and Biochemistry and
Rho, a member of the Rho small G protein family, regulates the
formation of stress fibers and focal adhesions in various types of cultured cells. We investigated here the actions of ROCK and mDia,
both of which have been identified to be putative downstream target
molecules of Rho, in Madin-Darby canine kidney cells. The dominant active mutant of RhoA induced the formation of parallel stress
fibers and focal adhesions, whereas the dominant active mutant of ROCK
induced the formation of stellate stress fibers and focal adhesions,
and the dominant active mutant of mDia induced the weak formation of
parallel stress fibers without affecting the formation of focal
adhesions. In the presence of C3 ADP-ribosyltransferase for Rho, the
dominant active mutant of ROCK induced the formation of stellate stress
fibers and focal adhesions, whereas the dominant active mutant of mDia
induced only the diffuse localization of actin filaments. These results
indicate that ROCK and mDia show distinct actions in reorganization of
the actin cytoskeleton. The dominant negative mutant of either ROCK or
mDia inhibited the formation of stress fibers and focal adhesions,
indicating that both ROCK and mDia are necessary for the formation of
stress fibers and focal adhesions. Moreover, inactivation and
reactivation of both ROCK and mDia were necessary for the
12-O-tetradecanoylphorbol-13-acetate-induced disassembly and reassembly, respectively, of stress fibers and focal
adhesions. The morphologies of stress fibers and focal adhesions in the
cells expressing both the dominant active mutants of ROCK and mDia were
not identical to those induced by the dominant active mutant of Rho.
These results indicate that at least ROCK and mDia cooperatively act as
downstream target molecules of Rho in the Rho-induced reorganization of
the actin cytoskeleton.
Second Department of Surgery, Osaka University Medical
School, Suita 565-0871, Japan
Corresponding author. E-mail address:
ytakai{at}molbio.med.osaka-u.ac.jp.
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