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J Biol Chem, Vol. 274, Issue 6, 3396-3401, February 5, 1999
,
From the Departments of RhoA has been identified as an important
regulator of cell proliferation. We recently showed that the Ras/RhoA
pathway regulates the degradation of p27Kip and the
progression of Chinese hamster embryo fibroblasts (IIC9 cells) through
G1 into S phase (Weber, J. D., Hu, W., Jefcoat, S. C.,
Raben, D. M., and Baldassare, J. J. (1997) J. Biol. Chem. 272, 32966-32971). In this report, we have demonstrated that, in
IIC9 cells, RhoA regulates cyclin E/CDK2 activity, which is required
for p27Kip degradation. As previously shown in several
fibroblasts cell lines, expression of dominant-negative CDK2 in IIC9
cells blocked serum-induced cyclin E/CDK2 activity and
p27Kip degradation. In the absence of serum, expression of
constitutively active RhoA(63) resulted in significant stimulation of
cyclin E/CDK2 activity and degradation of p27Kip.
Cotransfection of dominant-negative CDK2 and RhoA(63) inhibited RhoA(63)-induced cyclin E/CDK2 activity and p27Kip
degradation. In addition, expression of dominant-negative RhoA blocked
serum-induced cyclin E/CDK2 activity and p27Kip
degradation. Finally, expression of catalytically active cyclin E/CDK2
rescued the effect of expression of dominant-negative RhoA. Taken
together, these data show that RhoA regulates p27Kip
degradation through its regulation of cyclin E/CDK2 activity.
Pharmacological and
Physiological Science and § Molecular Microbiology and
Immunology, Saint Louis University School of Medicine,
St. Louis, Missouri 63110
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