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J. Biol. Chem., Vol. 278, Issue 39, 37865-37873, September 26, 2003

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Degradation of Cyclin B Is Required for the Onset of Anaphase in Mammalian Cells^*

Donald C. Chang   ¶, Naihan Xu  and Kathy Q. Luo 

From the Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China and the Marine Biological Laboratory, Woods Hole, Massachusetts 02543

Recently, it has been shown that cyclin B1 was degraded mainly before the onset of anaphase in mammalian cells. When a nondegradable form of cyclin B1 was introduced into cells, the metaphase-anaphase transition was blocked. This blockage was not due to a failure in activating anaphase-promoting complex, nor was it due to a failure of degradation of securin. To resolve the question of whether this blockage by overexpressing the nondegradable form of cyclin B1 is physiologically relevant or not, we developed a novel method to estimate the relative protein level of the overexpressed cyclin B1 mutant within an individual cell. We found that a low level of nondegradable cyclin B1 (less than 30% of the endogenous cyclin B1) was sufficient to block the metaphase-anaphase transition, implying that the blockage of anaphase onset by the nondegradable cyclin B1 was not due to an artifact of excessive M-phase-promoting factor activity. This result suggests that, in mammalian cells, the majority of cyclin B1 must be destroyed before the cell can enter anaphase.

Received for publication, June 17, 2003 , and in revised form, July 14, 2003.

^* This work was supported by the Research Grants Council of Hong Kong (HKUST6109/01 M and HKUST6104/02M) and the HIA project of HKUST. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed. Tel.: 852-2358-7326; Fax: 852-2358-1559; E-mail: bochang{at}ust.hk.

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