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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, 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]
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“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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Citations
Cites
Coot : model-building tools for molecular graphics
Emsley, P., & Cowtan, K. (2004) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444904019158 · Cited by 30,571
Jalview Version 2—a multiple sequence alignment editor and analysis workbench
Waterhouse, A. M., Procter, J. B., Martin, D. M. A., Clamp, M. E., & Barton, G. J. (2009) · Bioinformatics · doi:10.1093/bioinformatics/btp033 · Cited by 10,349
REFMAC 5 for the refinement of macromolecular crystal structures
Murshudov, G. N., Skubák, P., Lebedev, A. A., Pannu, N. S., Steiner, R. A., Nicholls, R. A., Winn, M., Long, F., & Vagin, A. A. (2011) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444911001314 · Cited by 8,415
Perkins, D. N., Pappin, D., Creasy, D. M., & Cottrell, J. S. (1999) · Electrophoresis · doi:10.1002/(sici)1522-2683(19991201)20:18 · Cited by 8,264
RELION: Implementation of a Bayesian approach to cryo-EM structure determination
Scheres, S. H. W. (2012) · Journal of Structural Biology · doi:10.1016/j.jsb.2012.09.006 · Cited by 5,446
Keller, A., Nesvizhskii, A., Kolker, E., & Aebersold, R. (2002) · Analytical Chemistry · doi:10.1021/ac025747h · Cited by 4,970
Gctf: Real-time CTF determination and correction
Zhang, K. (2015) · Journal of Structural Biology · doi:10.1016/j.jsb.2015.11.003 · Cited by 3,417
Li, X.-M., Mooney, P., Zheng, S., Booth, C., Braunfeld, M. B., Gubbens, S., Agard, D., & Cheng, Y.-F. (2013) · Nature Methods · doi:10.1038/nmeth.2472 · Cited by 1,866
Accurate determination of local defocus and specimen tilt in electron microscopy
Mindell, J. A., & Grigorieff, N. (2003) · Journal of Structural Biology · doi:10.1016/s1047-8477(03)00069-8 · Cited by 1,414
UCSF Chimera, MODELLER, and IMP: An integrated modeling system
Yang, Z. R., Lasker, K., Schneidman‐Duhovny, D., Webb, B., Huang, C. C., Pettersen, E. F., Goddard, T. D., Meng, E. C., Šali, A., & Ferrin, T. E. (2011) · Journal of Structural Biology · doi:10.1016/j.jsb.2011.09.006 · Cited by 637
Control of Cyclin Ubiquitination by CDK-Regulated Binding of Hct1 to the Anaphase Promoting Complex
Zachariae, W., Schwab, M., Nasmyth, K., & Seufert, W. (1998) · Science · doi:10.1126/science.282.5394.1721 · Cited by 511
Mitotic Regulation of the APC Activator Proteins CDC20 and CDH1
Kramer, E. R., Scheuringer, N., Podtelejnikov, A. V., Mann, M., & Peters, J. (2000) · Molecular Biology of the Cell · doi:10.1091/mbc.11.5.1555 · Cited by 455
Ribosome structures to near-atomic resolution from thirty thousand cryo-EM particles
Bai, X., Fernández, I. S., McMullan, G., & Scheres, S. H. W. (2013) · eLife · doi:10.7554/elife.00461 · Cited by 431
Mitotic regulation of the human anaphase‐promoting complex by phosphorylation
Kraft, C., Herzog, F., Gieffers, C., Mechtler, K., Hagting, A., Pines, J., & Peters, J. (2003) · The EMBO Journal · doi:10.1093/emboj/cdg627 · Cited by 421
Jaspersen, S. L., Charles, J. F., & Morgan, D. O. (1999) · Current Biology · doi:10.1016/s0960-9822(99)80111-0 · Cited by 408
Cubism and the cell cycle: the many faces of the APC/C
Pines, J. (2011) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3132 · Cited by 390
Semi-automated selection of cryo-EM particles in RELION-1.3
Scheres, S. (2015) · Journal of Structural Biology · doi:10.1016/j.jsb.2014.11.010 · Cited by 380
Beam-induced motion correction for sub-megadalton cryo-EM particles
Scheres, S. H. W. (2014) · eLife · doi:10.7554/elife.03665 · Cited by 341
Zeng, X., Sigoillot, F. D., Gaur, S. K., Choi, S., Pfaff, K. L., Oh, D., Hathaway, N. A., Dimova, N. V., Cuny, G. D., & King, R. W. (2010) · Cancer Cell · doi:10.1016/j.ccr.2010.08.010 · Cited by 332
Phosphorylation by Cdc28 Activates the Cdc20-Dependent Activity of the Anaphase-Promoting Complex
Rudner, A. D., & Murray, A. W. (2000) · The Journal of Cell Biology · doi:10.1083/jcb.149.7.1377 · Cited by 296
Initiation of Translation by Cricket Paralysis Virus IRES Requires Its Translocation in the Ribosome
Fernández, I. S., Bai, X., Murshudov, G. N., Scheres, S. H. W., & Ramakrishnan, V. (2014) · Cell · doi:10.1016/j.cell.2014.04.015 · Cited by 254
Atomic structure of the APC/C and its mechanism of protein ubiquitination
Chang, L., Zhang, Z., Yang, J., McLaughlin, S. H., & Barford, D. (2015) · Nature · doi:10.1038/nature14471 · Cited by 247
Self-citationStructure of the Anaphase-Promoting Complex/Cyclosome Interacting with a Mitotic Checkpoint Complex
Herzog, F., Primorac, I., Dube, P., Lénárt, P., Sander, B. O., Mechtler, K., Stark, H., & Peters, J. (2009) · Science · doi:10.1126/science.1163300 · Cited by 238
Molecular architecture and mechanism of the anaphase-promoting complex
Chang, L., Zhang, Z., Yang, J., McLaughlin, S. H., & Barford, D. (2014) · Nature · doi:10.1038/nature13543 · Cited by 212
Self-citationPanta rhei: The APC/C at steady state
Primorac, I., & Musacchio, A. (2013) · The Journal of Cell Biology · doi:10.1083/jcb.201301130 · Cited by 211
Yeast Hct1 recognizes the mitotic cyclin Clb2 and other substrates of the ubiquitin ligase APC
Schwab, M. G., Neutzner, M., Möcker, D., & Seufert, W. (2001) · The EMBO Journal · doi:10.1093/emboj/20.18.5165 · Cited by 209
TPR Subunits of the Anaphase-Promoting Complex Mediate Binding to the Activator Protein CDH1
Vodermaier, H. C., Gieffers, C., Maurer‐Stroh, S., Eisenhaber, F., & Peters, J. (2003) · Current Biology · doi:10.1016/s0960-9822(03)00581-5 · Cited by 208
Passmore, L. A., McCormack, E. A., Au, S. W. N., Paul, A., Willison, K. R., Harper, J. W., & Barford, D. (2003) · The EMBO Journal · doi:10.1093/emboj/cdg084 · Cited by 201
Self-citationReversible phosphorylation controls the activity of cyclosome-associated cyclin-ubiquitin ligase.
Lahav‐Baratz, S., Sudakin, V., RUDERMAN, J. V., & Hershko, A. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.20.9303 · Cited by 197
Golan, A., Yudkovsky, Y., & Hershko, A. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m111476200 · Cited by 173
Kimata, Y., Baxter, J. E., Fry, A. M., & Yamano, H. (2008) · Molecular Cell · doi:10.1016/j.molcel.2008.09.023 · Cited by 149
Patra, D., & Dunphy, W. G. (1998) · Genes & Development · doi:10.1101/gad.12.16.2549 · Cited by 142
Phosphorylation of the Cyclosome Is Required for Its Stimulation by Fizzy/cdc20
Shteinberg, M., Protopopov, Y., Listovsky, T., Brandeis, M., & Hershko, A. (1999) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1999.0884 · Cited by 132
Insights into the anaphase-promoting complex: a molecular machine that regulates mitosis
Chang, L., & Barford, D. (2014) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2014.08.003 · Cited by 106
Self-citationStructural analysis of human Cdc20 supports multisite degron recognition by APC/C
Tian, W., Li, B., Warrington, R. T., Tomchick, D. R., Yu, H., & Luo, X. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1213438109 · Cited by 106
Systematic Phosphorylation Analysis of Human Mitotic Protein Complexes
Hegemann, B., Hutchins, J. R. A., Hudecz, O., Novatchkova, M., Rameseder, J., Sykora, M. M., Liu, S., Mazanek, M., Lénárt, P., Hèriché, J.-K., et al. (2011) · Science Signaling · doi:10.1126/scisignal.2001993 · Cited by 104
Steen, J. A. J., Steen, H., Georgi, A. B., Parker, K. C., Springer, M., Kirchner, M., Hamprecht, F. A., & Kirschner, M. W. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0709807104 · Cited by 88
Mad2 and the APC/C compete for the same site on Cdc20 to ensure proper chromosome segregation
Izawa, D., & Pines, J. (2012) · The Journal of Cell Biology · doi:10.1083/jcb.201205170 · Cited by 86
van Zon, W., Ogink, J., ter Riet, B., Medema, R. H., Riele, H. T., & Wolthuis, R. M. F. (2010) · The Journal of Cell Biology · doi:10.1083/jcb.200912084 · Cited by 60
Recombinant expression, reconstitution and structure of human anaphase-promoting complex (APC/C)
Zhang, Z., Yang, J., Kong, E. H., Chao, W. C. H., Morris, E. P., da Fonseca, P. C. A., & Barford, D. (2012) · Biochemical Journal · doi:10.1042/bj20121374 · Cited by 59
Self-citationBinding of Activated Cyclosome to p13
Sudakin, V., Shteinberg, M., Ganoth, D., Hershko, J., & Hershko, A. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.29.18051 · Cited by 55
Activation of the APC/C Ubiquitin Ligase by Enhanced E2 Efficiency
Voorhis, V. A. V., & Morgan, D. O. (2014) · Current Biology · doi:10.1016/j.cub.2014.05.052 · Cited by 43
Yamaguchi, M., Yu, S., Qiao, R., Weissmann, F., Miller, D. J., VanderLinden, R. T., Brown, N. G., Frye, J. J., Peters, J., & Schulman, B. A. (2014) · Journal of Molecular Biology · doi:10.1016/j.jmb.2014.11.020 · Cited by 42
Role of Suc1 in the Activation of the Cyclosome by Protein Kinase Cdk1/Cyclin B
Shteinberg, M., & Hershko, A. (1999) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1999.0409 · Cited by 40
Herzog, F., Mechtler, K., & Peters, J. (2005) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(05)98019-1 · Cited by 18
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Targeted protein degradation: expanding the toolbox
Schapira, M., Calabrese, M. F., Bullock, A. N., & Crews, C. M. (2019) · Nature Reviews Drug Discovery · doi:10.1038/s41573-019-0047-y · Cited by 960
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Metadata
- 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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