Molecular mechanism of APC/C activation by mitotic phosphorylation
Open accessAbstract
In eukaryotes, the anaphase-promoting complex (APC/C, also known as the cyclosome) regulates the ubiquitin-dependent proteolysis of specific cell-cycle proteins to coordinate chromosome segregation in mitosis and entry into the G1 phase. The catalytic activity of the APC/C and its ability to specify the destruction of particular proteins at different phases of the cell cycle are controlled by its interaction with two structurally related coactivator subunits, Cdc20 and Cdh1. Coactivators recognize substrate degrons, and enhance the affinity of the APC/C for its cognate E2 (refs 4-6). During mitosis, cyclin-dependent kinase (Cdk) and polo-like kinase (Plk) control Cdc20- and Cdh1-mediated activation of the APC/C. Hyperphosphorylation of APC/C subunits, notably Apc1 and Apc3, is required for Cdc20 to activate the APC/C, whereas phosphorylation of Cdh1 prevents its association with the APC/C. Since both coactivators associate with the APC/C through their common C-box and Ile-Arg tail motifs, the mechanism underlying this differential regulation is unclear, as is the role of specific APC/C phosphorylation sites. Here, using cryo-electron microscopy and biochemical analysis, we define the molecular basis of how phosphorylation of human APC/C allows for its control by Cdc20. An auto-inhibitory segment of Apc1 acts as a molecular switch that in apo unphosphorylated APC/C interacts with the C-box binding site and obstructs engagement of Cdc20. Phosphorylation of the auto-inhibitory segment displaces it from the C-box-binding site. Efficient phosphorylation of the auto-inhibitory segment, and thus relief of auto-inhibition, requires the recruitment of Cdk-cyclin in complex with a Cdk regulatory subunit (Cks) to a hyperphosphorylated loop of Apc3. We also find that the small-molecule inhibitor, tosyl-l-arginine methyl ester, preferentially suppresses APC/C(Cdc20) rather than APC/C(Cdh1), and interacts with the binding sites of both the C-box and Ile-Arg tail motifs. Our results reveal the mechanism for the regulation of mitotic APC/C by phosphorylation and provide a rationale for the development of selective inhibitors of this state.
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CDC20 uses the side surface of its WDR domain to recruit APC substrates via their D-box motif.
“CDC20, a non-DCAF E3 that acts as the substratebinding subunit of the APC, uses the side surface of its WDR domain to recruit APC substrates via their D-box motif 73”
From Targeted protein degradation: expanding the toolbox (2019) · cites this paper as [73]
The structure of the CDC20 WDR domain was solved in complex with a substrate peptide in which a canonical RxxL D-box motif is inserted into a hydrophobic cavity on the side surface of the WDR domain.
“The structure of the CDC20 WDR domain was solved in complex with a substrate peptide in which a canonical RxxL D-box motif is inserted into a hydrophobic cavity on the side surface of the WDR domain 73 .”
From Targeted protein degradation: expanding the toolbox (2019) · cites this paper as [73]
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- Authors
- Zhang, S., Chang, L., Alfieri, C., Zhang, Z., Yang, J., Maslen, S., Skehel, M., & Barford, D.
- Affiliations
- MRC Laboratory of Molecular Biology (GB)
- Corresponding author
- Barford, David
- ORCID
- Venue
- Nature
- Published
- 2016-04-27
- Volume
- 533
- Issue
- 7602
- Pages
- 260-264
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4878669
- OpenAlex
- W2343854987
- Cited by
- 203
- References
- 45
- Citations by year
- 2026: 6 · 2025: 14 · 2024: 22 · 2023: 20 · 2022: 15 · 2021: 21 · 2020: 26 · 2019: 26 · 2018: 16 · 2017: 26 · 2016: 10
- FWCI
- 17.91
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Cancer Research UK (14109)
- Boehringer Ingelheim Fonds
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Keywords
- anaphase-promoting complex, mitotic phosphorylation, CDC20, CDH1, E3 ubiquitin ligase, cyclin-dependent kinases, Polo-like kinase, cell cycle regulation, chromosome segregation, ubiquitin-mediated proteolysis, cryo-electron microscopy
- MeSH terms
- Apoenzymes, Binding Sites, Enzyme Activation, Humans, Mitosis, Models, Molecular, Phosphoproteins, Phosphorylation, Protein Binding, Protein Conformation, Tosylarginine Methyl Ester, Antigens, CD, Cadherins, Cyclins, Cyclin-Dependent Kinases, Cryoelectron Microscopy, Amino Acid Motifs, Anaphase-Promoting Complex-Cyclosome, Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome, Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome, Cdc20 Proteins
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