The spindle checkpoint requires cyclin-dependent kinase activity

The spindle checkpoint prevents anaphase onset until completion of mitotic spindle assembly by restraining activation of the ubiquitin ligase anaphase-promoting complex/cyclosome-Cdc20 (APC/C-Cdc20). We show that the spindle checkpoint requires mitotic cyclin-dependent kinase (cdk) activity. Inhibiting cdk activity overrides checkpoint-dependent arrest in Xenopus egg extracts and human cells. Following inhibition, the interaction between APC/C and Cdc20 transiently increases while the inhibitory checkpoint protein Mad2 dissociates from Cdc20. Cdk inhibition also overcomes Mad2-induced mitotic arrest. in addition, in vitro cdk1-phosphorylated Cdc20 interacts with Mad2 rather than APC/C. Thus, cdk activity is required to restrain APC/C-Cdc20 activation until completion of spindle assembly.