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Originally published as MBC in Press, 10.1091/mbc.E07-03-0281 on July 18, 2007

Vol. 18, Issue 10, 3752-3763, October 2007

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Inactivation of Rho GTPases with Clostridium difficile Toxin B Impairs Centrosomal Activation of Aurora-A in G2/M Transition of HeLa Cells

Yoshikazu Ando*, Shingo Yasuda*,{dagger}, Fabian Oceguera-Yanez*, and Shuh Narumiya*

*Department of Pharmacology and {dagger}Horizontal Medical Research Organization, Kyoto University Faculty of Medicine, Kyoto 606-8501, Japan

Submitted March 27, 2007; Revised June 29, 2007; Accepted July 6, 2007
Monitoring Editor: Yixian Zheng

During G2 phase of cell cycle, centrosomes function as a scaffold for activation of mitotic kinases. Aurora-A is first activated at late G2 phase at the centrosome, facilitates centrosome maturation, and induces activation of cyclin B-Cdk1 at the centrosome for mitotic entry. Although several molecules including HEF1 and PAK are implicated in centrosomal activation of Aurora-A, signaling pathways leading to Aurora-A activation at the centrosome, and hence mitotic commitment in vertebrate cells remains largely unknown. Here, we have used Clostridium difficile toxin B and examined the role of Rho GTPases in G2/M transition of HeLa cells. Inactivation of Rho GTPases by the toxin B treatment delayed by 2 h histone H3 phosphorylation, Cdk1/cyclin B activation, and Aurora-A activation. Furthermore, PAK activation at the centrosome that was already present before the toxin addition was significantly attenuated for 2 h by the addition of toxin B, and HEF1 accumulation at the centrosome that occurred in late G2 phase was also delayed. These results suggest that Rho GTPases function in G2/M transition of mammalian cells by mediating multiple signaling pathways converging to centrosomal activation of Aurora-A.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E07-03-0281) on July 18, 2007.

Address correspondence to: Shuh Narumiya (snaru{at}mfour.med.kyoto-u.ac.jp)




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