p31^sup comet^ promotes disassembly of the mitotic checkpoint complex in an ATP-dependent process

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 108; no. 8; p. 3187
• Main Authors: Teichner, Adar, Eytan, Esther, Sitry-Shevah, Danielle, Miniowitz-Shemtov, Shirly, Dumin, Elena, Gromis, Jonathan, Hershko, Avram
• Format: Journal Article
• Language: English
• Published: Washington National Academy of Sciences 22.02.2011
• Subjects: Adenosine triphosphatase; Biochemistry; Cell cycle; Cell division; Chromatin; Chromosomes
• ISSN: 0027-8424; 1091-6490

Accurate segregation of chromosomes in mitosis is ensured by a surveillance mechanism called the mitotic (or spindle assembly) checkpoint. It prevents sister chromatid separation until all chromosomes are correctly attached to the mitotic spindle through their kinetochores. The checkpoint acts by inhibiting the anaphase-promoting complex/cyclosome (APC/C), a ubiquitin ligase that targets for degradation securin, an inhibitor of anaphase initiation. The activity of APC/C is inhibited by a mitotic checkpoint complex (MCC), composed of the APC/C activator Cdc20 bound to the checkpoint proteins MAD2, BubR1, and Bub3. When all kinetochores acquire bipolar attachment the checkpoint is inactivated, but the mechanisms of checkpoint inactivation are not understood. We have previously observed that hydrolyzable ATP is required for exit from checkpoint-arrested state. In this investigation we examined the possibility that ATP hydrolysis in exit from checkpoint is linked to the action of the Mad2-binding protein p31... in this process. It is known that p31... prevents the formation of a Mad2 dimer that it thought to be important for turning on the mitotic checkpoint. This explains how p31... blocks the activation of the checkpoint but not how it promotes its inactivation. Using extracts from checkpoint-arrested cells and MCC isolated from such extracts, we now show that p31... causes the disassembly of MCC and that this process requires β,...-hydrolyzable ATP. Although p31... binds to Mad2, it promotes the dissociation of Cdc20 from BubR1 in MCC. (ProQuest: ... denotes formulae/symbols omitted.)