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Published 24 June 2002. doi:10.1083/jcb.200203035
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© The Rockefeller University Press, 0021-9525/2002/6/1139 $5.00
The Journal of Cell Biology, Volume 157, Number 7, June 24, 2002 1139-1149

Article

 The roles of Fzy/Cdc20 and Fzr/Cdh1 in regulating the destruction of cyclin B in space and time

Jordan W. Raff, Kim Jeffers and Jun-yong Huang

Department of Genetics, Wellcome/Cancer Research UK Institute, Cambridge CB2 1QR, UK

Address correspondence to Jordan W. Raff, Department of Genetics, Wellcome/Cancer Research UK Institute, Cambridge CB2 1QR, UK. Tel.: 44-122-333-4114. Fax: 44-122-333-4089. E-mail: j.raff{at}welc.cam.ac.uk

In Drosophila cells cyclin B is normally degraded in two phases: (a) destruction of the spindle-associated cyclin B initiates at centrosomes and spreads to the spindle equator; and (b) any remaining cytoplasmic cyclin B is degraded slightly later in mitosis. We show that the APC/C regulators Fizzy (Fzy)/Cdc20 and Fzy-related (Fzr)/Cdh1 bind to microtubules in vitro and associate with spindles in vivo. Fzy/Cdc20 is concentrated at kinetochores and centrosomes early in mitosis, whereas Fzr/Cdh1 is concentrated at centrosomes throughout the cell cycle. In syncytial embryos, only Fzy/Cdc20 is present, and only the spindle-associated cyclin B is degraded at the end of mitosis. A destruction box–mutated form of cyclin B (cyclin B triple-point mutant [CBTPM]–GFP) that cannot be targeted for destruction by Fzy/Cdc20, is no longer degraded on spindles in syncytial embryos. However, CBTPM–GFP can be targeted for destruction by Fzr/Cdh1. In cellularized embryos, which normally express Fzr/Cdh1, CBTPM–GFP is degraded throughout the cell but with slowed kinetics. These findings suggest that Fzy/Cdc20 is responsible for catalyzing the first phase of cyclin B destruction that occurs on the mitotic spindle, whereas Fzr/Cdh1 is responsible for catalyzing the second phase of cyclin B destruction that occurs throughout the cell. These observations have important implications for the mechanisms of the spindle checkpoint.

Key Words: Cdc20; Cdh1; anaphase-promoting complex; cyclin B; mitosis


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