Unattached Kinetochores Catalyze Production of an Anaphase Inhibitor that Requires a Mad2 Template to Prime Cdc20 for BubR1 Binding
Open accessAbstract
Premature anaphase onset is prevented by the mitotic checkpoint through production of a "wait anaphase" inhibitor(s) that blocks recognition of cyclin B and securin by Cdc20-activated APC/C, an E3 ubiquitin ligase that targets them for destruction. Using physiologically relevant levels of Mad2, Bub3, BubR1, and Cdc20, we demonstrate that unattached kinetochores on purified chromosomes catalytically generate a diffusible Cdc20 inhibitor or inhibit Cdc20 already bound to APC/C. Furthermore, the chromosome-produced inhibitor requires both recruitment of Mad2 by Mad1 that is stably bound at unattached kinetochores and dimerization-competent Mad2. We show that purified chromosomes promote BubR1 binding to APC/C-Cdc20 by acting directly on Mad2, but not BubR1. Our results support a model in which immobilized Mad1/Mad2 at kinetochores provides a template for initial assembly of Mad2 bound to Cdc20 that is then converted to a final mitotic checkpoint inhibitor with Cdc20 bound to BubR1.
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- Authors
- Kulukian, A., Han, J. S., & Cleveland, D. W.
- Affiliations
- University of California San Diego (US)
- Corresponding author
- Cleveland, Don W.
- Venue
- Developmental Cell
- Publisher
- Elsevier BV
- Published
- 2009-01
- Volume
- 16
- Issue
- 1
- Pages
- 105-117
- ISSN
- 1534-5807 (ISSN-L)
- PMC
- PMC2655205
- OpenAlex
- W2139141152
- Cited by
- 246
- References
- 55
- Citations by year
- 2026: 1 · 2025: 9 · 2024: 7 · 2023: 7 · 2022: 8 · 2021: 7 · 2020: 7 · 2019: 10 · 2018: 10 · 2017: 10 · 2016: 12 · 2015: 19 · 2014: 24 · 2013: 20 · 2012: 26
- FWCI
- 14.09
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Developmental Cell (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (R37GM029513, GM 29513, R01GM029513)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- spindle assembly checkpoint, Mad2, Mad1, BubR1, CDC20, Bub3, anaphase-promoting complex, securin, cyclin B, ubiquitin ligase, kinetochore-microtubule attachment
- MeSH terms
- Poly-ADP-Ribose Binding Proteins, Anaphase, Calcium-Binding Proteins, Chromosomes, HeLa Cells, Humans, Nuclear Proteins, Recombinant Proteins, Repressor Proteins, Protein Serine-Threonine Kinases, Kinetochores, Cell Cycle Proteins, Dimerization, Ubiquitin-Protein Ligase Complexes, Anaphase-Promoting Complex-Cyclosome, Cdc20 Proteins, Mad2 Proteins
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