TY - JOUR
T1 - RhoA GTPase is dispensable for actomyosin regulation but is essential for mitosis in primary Mouse embryonic fibroblasts
AU - Melendez, Jaime
AU - Stengel, Kristy
AU - Zhou, Xuan
AU - Chauhan, Bharesh K.
AU - Debidda, Marcella
AU - Andreassen, Paul
AU - Lang, Richard A.
AU - Zheng, Yi
PY - 2011/4/29
Y1 - 2011/4/29
N2 - RhoA, the founding member of mammalian Rho GTPase family, is thought to be essential for actomyosin regulation. To date, the physiologic function of RhoA in mammalian cell regulation has yet to be determined genetically. Here we have created RhoA conditional knock-out mice. Mouse embryonic fibroblasts deleted of RhoA showed no significant change in actin stress fiber or focal adhesion complex formation in response to serum or LPA, nor any detectable change in Rho-kinase signaling activity. Concomitant knock-out or knockdown of RhoB and RhoC in the RhoA-/- cells resulted in a loss of actin stress fiber and focal adhesion similar to that of C3 toxin treatment. Proliferation of RhoA-/- cells was impaired due to a complete cell cycle block during mitosis, an effect that is associated with defective cytokinesis and chromosome segregation and can be readily rescued by exogenous expression of RhoA. Furthermore, RhoA deletion did not affect the transcriptional activity of Stat3, NFκB, or serum response factor, nor the expression of the cell division kinase inhibitor p21Cip1 or p27Kip1. These genetic results demonstrate that in primary mouse embryonic fibroblasts, RhoA is uniquely required for cell mitosis but is redundant with related RhoB and RhoC GTPases in actomyosin regulation.
AB - RhoA, the founding member of mammalian Rho GTPase family, is thought to be essential for actomyosin regulation. To date, the physiologic function of RhoA in mammalian cell regulation has yet to be determined genetically. Here we have created RhoA conditional knock-out mice. Mouse embryonic fibroblasts deleted of RhoA showed no significant change in actin stress fiber or focal adhesion complex formation in response to serum or LPA, nor any detectable change in Rho-kinase signaling activity. Concomitant knock-out or knockdown of RhoB and RhoC in the RhoA-/- cells resulted in a loss of actin stress fiber and focal adhesion similar to that of C3 toxin treatment. Proliferation of RhoA-/- cells was impaired due to a complete cell cycle block during mitosis, an effect that is associated with defective cytokinesis and chromosome segregation and can be readily rescued by exogenous expression of RhoA. Furthermore, RhoA deletion did not affect the transcriptional activity of Stat3, NFκB, or serum response factor, nor the expression of the cell division kinase inhibitor p21Cip1 or p27Kip1. These genetic results demonstrate that in primary mouse embryonic fibroblasts, RhoA is uniquely required for cell mitosis but is redundant with related RhoB and RhoC GTPases in actomyosin regulation.
UR - http://www.scopus.com/inward/record.url?scp=79955382980&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79955382980&partnerID=8YFLogxK
U2 - 10.1074/jbc.C111.229336
DO - 10.1074/jbc.C111.229336
M3 - Article
C2 - 21454503
AN - SCOPUS:79955382980
SN - 0021-9258
VL - 286
SP - 15132
EP - 15137
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 17
ER -