Inhibitory factors associated with anaphase-promoting complex/cylosome in mitotic checkpoint

Braunstein, Ilana; Miniowitz, Shirly; Moshe, Yakir; Hershko, Avram

doi:10.1073/pnas.0700523104

Cited by 70 publications

(97 citation statements)

References 35 publications

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“…When such extracts were incubated in the presence of ATP, APC/C was converted to an active form following a lag period. This process was accompanied by the release of inhibitory factors from APC/C and the disassembly of MCC (8,9). We found that the hydrolysis of ATP at the β−γ position was required for both for the activation of APC/C and for the disassembly of MCC (10); this was different from the previously described requirement of checkpoint inactivation for ubiquitylation (11,12), because the latter process involves the cleavage of the α−β bond of ATP (13).…”

contrasting

confidence: 53%

“…Extracts from nocodazole-arrested HeLa cells were prepared as described previously (9). To isolate MCC free of APC/C, extracts were subjected to sequential immunoprecipitations with anti-Cdc27 followed by anti-BubR1 antibodies [both bound to Affi-Prep Protein A beads (Bio-Rad)], as described (10,17).…”

Section: Methodsmentioning

confidence: 99%

“…For this purpose, we used extracts from nocodazole-arrested cells that faithfully reproduced downstream events of the mitotic checkpoint system (9). When such extracts were incubated in the presence of ATP, APC/C was converted to an active form following a lag period.…”

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confidence: 99%

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Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Miniowitz-Shemtov

Eytan

Ganoth

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The mitotic checkpoint system delays anaphase until all chromosomes are correctly attached to the mitotic spindle. When the checkpoint is turned on, it promotes the formation of the mitotic checkpoint complex (MCC), which inhibits the ubiquitin ligase anaphase-promoting complex/cyclosome (APC/C). MCC is composed of the checkpoint proteins BubR1, Bub3, and Mad2 bound to the APC/C activator Cdc20. When the checkpoint is satisfied, MCC is disassembled and APC/C becomes active. Previous studies have shown that the Mad2-binding protein p31 comet promotes the dissociation of Cdc20 from BubR1 in MCC in a process that requires ATP. We now show that a part of MCC dissociation is blocked by inhibitors of cyclin-dependent kinases (Cdks) and that purified Cdk1–cyclin B stimulates this process. The mutation of all eight potential Cdk phosphorylation sites of Cdc20 partially prevented its release from BubR1. Furthermore, p31 comet stimulated Cdk-catalyzed phosphorylation of Cdc20 in MCC. It is suggested that the binding of p31 comet to Mad2 in MCC may trigger a conformational change in Cdc20 that facilitates its phosphorylation by Cdk, and that the latter process may promote its dissociation from BubR1.

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confidence: 53%

Section: Methodsmentioning

confidence: 99%

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Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Miniowitz-Shemtov

Eytan

Ganoth

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…As shown in Fig. 4, in sham-depleted extract cyclin B was degraded after a lag, as described (9,23). In TRIP13, p31 comet double-depleted extract, there was little degradation of cyclin B after 3 h of incubation.…”

Section: Resultsmentioning

confidence: 91%

Disassembly of mitotic checkpoint complexes by the joint action of the AAA-ATPase TRIP13 and p31 ^comet

Eytan

Wang

Miniowitz-Shemtov

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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131

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Significance The mitotic checkpoint system has an important role to ensure accurate segregation of chromosomes in mitosis. This system regulates the activity of the ubiquitin ligase Anaphase-Promoting Complex/Cyclosome (APC/C) by the formation of a negatively acting Mitotic Checkpoint Complex (MCC). When the checkpoint is satisfied, MCC is disassembled, but the mechanisms of MCC disassembly are not well understood. We show here that the ATP-hydrolyzing enzyme Thyroid Receptor Interacting Protein 13 (TRIP13), along with the MCC-targeting protein p31 comet , promote the disassembly of the mitotic checkpoint complexes and the inactivation of the mitotic checkpoint. The results reveal an important molecular mechanism in the regulation of APC/C by the mitotic checkpoint.

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“…The MCC may also bind to APC in a kinetochore independent manner to eventually inhibit APC by releasing O-Mad2, forming the stable Bub3:BubR1:Cdc20:APC complex. The recently discovered mitotic checkpoint factor 2 protein (MCF2) is a highly potent APC inhibitor, yet the mechanism of binding to the APC and its regulation is still elusive (Braunstein et al, 2007;Eytan et al, 2008). All complexes inhibiting APC (except of MCF2) rely on the presence of Cdc20:C-Mad2, which requires unattached kinetochores for adequately fast formation.…”

Section: Apc Inhibitionmentioning

confidence: 99%

Systems Biology Modeling of Five Pathways for Regulation and Potent Inhibition of the Anaphase-Promoting Complex (APC/C): Pivotal Roles for MCC and BubR1

Ibrahim

2015

OMICS: A Journal of Integrative Biology

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Correct DNA segregation is a fundamental process that ensures the precise and reliable inheritance of genomic information for the propagation of cell life. Eukaryotic cells have evolved a conserved surveillance control mechanism for DNA segregation named the Spindle Assembly Checkpoint (SAC).The SAC ensures that the sister chromatids of the duplicated genome are not separated and distributed to the spindle poles before all chromosomes have been properly linked to the microtubules of the mitotic spindle. Biochemically, the SAC delays cell cycle progression by preventing activation of the anaphase-promoting complex (APC/C) or cyclosome whose activation by Cdc20 is required for sister-chromatid separation; this marks the transition into anaphase. In response to activation of the checkpoint, various species control the activity of both APC/C and Cdc20. However, the underlying regulatory pathways remain largely elusive. In this study, five possible model variants of APC/C regulation were constructed, namely BubR1, Mad2, MCC, MCF2, and an all-pathways model variant. These models were validated with experimental data from the literature. A wide range of parameter values has been tested to find the critical values of the APC/C binding rate. The results show that all variants are able to capture the wild-type behavior of the APC/C. However, only one model variant, which included both MCC as well as BubR1 as potent inhibitors of the APC/C, was able to reproduce both wild-type and mutant type behavior of APC/C regulation. In conclusion, the presented work informs the regulation of fundamental processes such as SAC and APC/C in cell biology and has successfully distinguished between five competing dynamical models using a systems biology approach. The results attest that systems-level approaches are vital for molecular and cell biology.

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Inhibitory factors associated with anaphase-promoting complex/cylosome in mitotic checkpoint

Cited by 70 publications

References 35 publications

Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Disassembly of mitotic checkpoint complexes by the joint action of the AAA-ATPase TRIP13 and p31 ^comet

Systems Biology Modeling of Five Pathways for Regulation and Potent Inhibition of the Anaphase-Promoting Complex (APC/C): Pivotal Roles for MCC and BubR1

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Inhibitory factors associated with anaphase-promoting complex/cylosome in mitotic checkpoint

Cited by 70 publications

References 35 publications

Role of phosphorylation of Cdc20 in p31 comet -stimulated disassembly of the mitotic checkpoint complex

Role of phosphorylation of Cdc20 in p31 comet -stimulated disassembly of the mitotic checkpoint complex

Disassembly of mitotic checkpoint complexes by the joint action of the AAA-ATPase TRIP13 and p31 comet

Systems Biology Modeling of Five Pathways for Regulation and Potent Inhibition of the Anaphase-Promoting Complex (APC/C): Pivotal Roles for MCC and BubR1

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Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Role of phosphorylation of Cdc20 in p31 ^comet -stimulated disassembly of the mitotic checkpoint complex

Disassembly of mitotic checkpoint complexes by the joint action of the AAA-ATPase TRIP13 and p31 ^comet