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J Biol Chem, Vol. 273, Issue 15, 8616-8622, April 10, 1998
Subunits and the Yeast
Response Regulator Protein Skn7
,
From the To identify potential RhoA effector proteins, we
conducted a two-hybrid screen for cDNAs encoding proteins that
interact with a Gal4-RhoA.V14 fusion protein. In addition to the RhoA
effector ROCK-I we identified cDNAs encoding Kinectin, mDia2 (a
p140 mDia-related protein), and the guanine nucleotide exchange factor,
mNET1. ROCK-I, Kinectin, and mDia2 can bind the wild type forms of both
RhoA and Cdc42 in a GTP-dependent manner in
vitro. Comparison of the ROCK-I and Kinectin sequences revealed a
short region of sequence homology that is both required for interaction
in the two-hybrid assay and sufficient for weak interaction in
vitro. Sequences related to the ROCK-I/Kinectin sequence homology
are present in heterotrimeric G protein Transcription Laboratory, Imperial Cancer
Research Fund Laboratories, 44 Lincoln's Inn Fields,
London WC2A 3PX and 
Division of Yeast Genetics, National
Institute for Medical Research, The Ridgeway, Mill Hill,
London NW7 1AA, United Kingdom
subunits and in the
Saccharomyces cerevisiae Skn7 protein. We show that 2
and Skn7 can interact with mammalian RhoA and Cdc42 and yeast Rho1,
both in vivo and in vitro. Functional assays in
yeast suggest that the Skn7 ROCK-I/Kinectin homology region is required
for its function in vivo.
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