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(Received for publication, December 27, 1996, and in revised form, August 25, 1997)
From the Division of Applied Toxicology, Institute of Toxicology,
University of Mainz, Obere Zahlbacher Str. 67, D-55131 Mainz, Germany
The small GTPase RhoB is immediate-early
inducible by DNA damaging treatments and thus part of the early
response of eukaryotic cells to genotoxic stress. To investigate the
regulation of this cellular response, we isolated the gene for
rhoB from a mouse genomic library. Sequence analysis of the
rhoB gene showed that its coding region does not contain
introns. The promoter region of rhoB harbors regulatory
elements such as TATA, CAAT, and Sp1 boxes but not consensus sequences
for AP-1, Elk-1, or c-Jun/ATF-2. The rhoB promoter was
activated by UV irradiation, but not by 12-O-tetradecanoylphorbol-13-acetate treatment.
rhoB promoter deletion constructs revealed a fragment of
0.17 kilobases in size which was sufficient in eliciting the UV
response. This minimal promoter fragment contains TATA and CAAT boxes
but no other known regulatory elements. Neither MEK inhibitor PD98059
nor p38 kinase inhibitor SB203580 blocked stimulation of
rhoB by UVC (UV light, 254 nm) which indicates that ERK or
p38 mitogen-activated protein (MAP) kinase are not involved in the UV
induction of rhoB. Also, phosphatidylinositol 3-kinase
inhibitor wortmannin, which blocks UV stimulation of both JNK and p38
MAP kinase, did not inhibit rhoB activation. Furthermore,
activation of JNK by interleukin-1
Volume 272, Number 49,
Issue of December 5, 1997
pp. 30637-30644
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
rhoB Encoding a UV-inducible Ras-related Small
GTP-binding Protein Is Regulated by GTPases of the Rho Family and
Independent of JNK, ERK, and p38 MAP Kinase
did not affect rhoB
expression. These data indicate that JNK is not involved in the
regulation of rhoB. Overexpression of wild-type Rac as well
as the Rho guanine-dissociation inhibitor caused activation of
rhoB. Wild-type RhoB inhibited both basal and UV-stimulated rhoB promoter activity, indicating a negative regulatory
feedback by RhoB itself. The data provide evidence both for a signal
transduction pathway independent of JNK, ERK, and p38 MAP kinase to be
involved in the induction of rhoB by genotoxic stress, and
furthermore, indicate autoregulation of rhoB.
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