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*Department of Molecular Biology and Biochemistry and
The Rho subfamily of the Rho small G protein family (Rho)
regulates formation of stress fibers and focal adhesions in many types
of cultured cells. In moving cells, dynamic and coordinate disassembly
and reassembly of stress fibers and focal adhesions are observed, but
the precise mechanisms in the regulation of these processes are poorly
understood. We previously showed that 12-O-tetradecanoylphorbol-13-acetate (TPA) first induced
disassembly of stress fibers and focal adhesions followed by their
reassembly in MDCK cells. The reassembled stress fibers showed
radial-like morphology that was apparently different from the original.
We analyzed here the mechanisms of these TPA-induced processes. Rho inactivation and activation were necessary for the TPA-induced disassembly and reassembly, respectively, of stress fibers and focal
adhesions. Both inactivation and activation of the Rac subfamily of the
Rho family (Rac) inhibited the TPA-induced reassembly of stress fibers
and focal adhesions but not their TPA-induced disassembly. Moreover,
microinjection or transient expression of Rab GDI, a regulator of all
the Rab small G protein family members, inhibited the TPA-induced
reassembly of stress fibers and focal adhesions but not their
TPA-induced disassembly, indicating that, furthermore, activation of
some Rab family members is necessary for their TPA-induced reassembly.
Of the Rab family members, at least Rab5 activation was necessary for
the TPA-induced reassembly of stress fibers and focal adhesions. The
TPA-induced, small G protein-mediated reorganization of stress fibers
and focal adhesions was closely related to the TPA-induced cell
motility. These results indicate that the Rho and Rab family members
coordinately regulate the TPA-induced reorganization of stress fibers
and focal adhesions that may cause cell motility.
Second Department of Surgery, Osaka University Medical
School, Suita 565-0871, Japan
Corresponding author.
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