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A more recent version of this article appeared on September 26, 2003
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M306376200v1
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Papers In Press, published online ahead of print July 14, 2003
J. Biol. Chem, 10.1074/jbc.M306376200
Submitted on June 17, 2003
Revised on July 14, 2003
Accepted on July 14, 2003

Degradation of cyclin B is required for the onset of anaphase in mammalian cells

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

Department of Biology, Hong Kong University of Science & Technology, Hong Kong, Hong Kong

Corresponding Author: bochang{at}ust.hk

Recently, it has been shown that cyclin B1 was degraded mainly before the onset of anaphase in mammalian cells. When nondegradable form of cyclin B1 was introduced into cells, the metaphase-anaphase (M/A) transition was blocked. This blockage was not due to a failure in activating APC (anaphase-promoting complex), neither was it due to a failure of degradation of securin. In order 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 M/A transition, implying that the blockage of anaphase onset by the nondegradable cyclin B1 was not due to an artifact of excessive MPF (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 anaphas


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