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PMID: 22940250 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

The APC/C subunit Mnd2/Apc15 promotes Cdc20 autoubiquitination and spindle assembly checkpoint inactivation.

Molecular cell ·Vol. 47 ·No. 6 ·2012-09-28 ·Pages 921-32

Foster SA, Morgan DO

Abstract

The fidelity of chromosome segregation depends on the spindle assembly checkpoint (SAC). In the presence of unattached kinetochores, anaphase is delayed when three SAC components (Mad2, Mad3/BubR1, and Bub3) inhibit Cdc20, the activating subunit of the anaphase-promoting complex (APC/C). We analyzed the role of Cdc20 autoubiquitination in the SAC of budding yeast. Reconstitution with purified components revealed that a Mad3-Bub3 complex synergizes with Mad2 to lock Cdc20 on the APC/C and stimulate Cdc20 autoubiquitination, while inhibiting ubiquitination of substrates. SAC-dependent Cdc20 autoubiquitination required the Mnd2/Apc15 subunit of the APC/C. General inhibition of Cdc20 ubiquitination in vivo resulted in high Cdc20 levels and a failure to establish a SAC arrest, suggesting that SAC establishment depends on low Cdc20 levels. Specific inhibition of SAC-dependent ubiquitination, by deletion of Mnd2, allowed establishment of a SAC arrest but delayed release from the arrest, suggesting that Cdc20 ubiquitination is also required for SAC inactivation.

MeSH Terms
Anaphase-Promoting Complex-Cyclosome Cdc20 Proteins Cell Cycle Proteins/genetics,metabolism Chromosome Segregation M Phase Cell Cycle Checkpoints Mad2 Proteins Nuclear Proteins/metabolism Saccharomyces cerevisiae Proteins/metabolism Spindle Apparatus/genetics,metabolism Ubiquitin-Protein Ligase Complexes/metabolism Ubiquitination
Chemicals
BUB3 protein, S cerevisiae CDC20 protein, S cerevisiae Cdc20 Proteins Cell Cycle Proteins MAD2 protein, S cerevisiae MAD3 protein, S cerevisiae Mad2 Proteins Nuclear Proteins Saccharomyces cerevisiae Proteins Ubiquitin-Protein Ligase Complexes Anaphase-Promoting Complex-Cyclosome
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Foster Scott A
Department of Physiology and Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158, USA.
Morgan David O
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2012.07.031
Published
2012-09-28
Epub
2012-00-30
Pages
921-32
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC3462283
Subset
IM
Grants
NIGMS NIH HHS · R01 GM053270 · United States
NIGMS NIH HHS · R37 GM053270 · United States
NIGMS NIH HHS · GM053270 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2012.07.031 PMC Full Text View journal details
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