Abstract
Aneuploidy and chromosomal instability are major characteristics of human cancer. These abnormalities can result from defects in the spindle assembly checkpoint (SAC), which is a surveillance mechanism for accurate chromosome segregation through restraint of the activity of the anaphase-promoting complex/cyclosome (APC/C). Here, we show that a CUE-domain-containing protein, CUEDC2, is a cell-cycle regulator that promotes spindle checkpoint inactivation and releases APC/C from checkpoint inhibition. CUEDC2 is phosphorylated by Cdk1 during mitosis. Depletion of CUEDC2 causes a checkpoint-dependent delay of the metaphase–anaphase transition. Phosphorylated CUEDC2 binds to Cdc20, an activator of APC/C, and promotes the release of Mad2 from APC/C–Cdc20 and subsequent APC/C activation. CUEDC2 overexpression causes earlier activation of APC/C, leading to chromosome missegregation and aneuploidy. Interestingly, CUEDC2 is highly expressed in many types of tumours. These results suggest that CUEDC2 is a key regulator of mitosis progression, and that CUEDC2 dysregulation might contribute to tumour development by causing chromosomal instability.
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References
Weaver, B. A. & Cleveland, D. W. The role of aneuploidy in promoting and suppressing tumors. J. Cell Biol. 185, 935–937 (2009).
Nigg, E. A. Mitotic kinases as regulators of cell division and its checkpoints. Nat. Rev. Mol. Cell Biol. 2, 21–32 (2001).
Pines, J. & Rieder, C. L. Re-staging mitosis: a contemporary view of mitotic progression. Nat. Cell Biol. 3, E3–E6 (2001).
Morgan, D. O. Regulation of the APC and the exit from mitosis. Nat. Cell Biol. 1, E47–E53 (1999).
Page, A. M. & Hieter, P. The anaphase-promoting complex: new subunits and regulators. Annu. Rev. Biochem. 68, 583–609 (1999).
Peters, J. M. The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat. Rev. Mol. Cell Biol. 7, 644–656 (2006).
Sullivan, M. & Morgan, D. O. Finishing mitosis, one step at a time. Nat. Rev. Mol. Cell Biol. 8, 894–903 (2007).
Visintin, R., Prinz, S. & Amon, A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 278, 460–463 (1997).
Hagting, A. et al. Human securin proteolysis is controlled by the spindle checkpoint and reveals when the APC/C switches from activation by Cdc20 to Cdh1. J. Cell Biol. 157, 1125–1137 (2002).
Visintin, R. et al. The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk-dependent phosphorylation. Mol. Cell 2, 709–718 (1998).
Nasmyth, K. Segregating sister genomes: the molecular biology of chromosome separation. Science 297, 559–565 (2002).
Murray, A. W. Recycling the cell cycle: cyclins revisited. Cell 116, 221–234 (2004).
Hershko, A. Mechanisms and regulation of the degradation of cyclin B. Philos. Trans. R. Soc. Lond. B Biol. Sci. 354, 1571–1575 (1999) discussion 1575–1576.
Glotzer, M., Murray, A. W. & Kirschner, M. W. Cyclin is degraded by the ubiquitin pathway. Nature 349, 132–138 (1991).
Bharadwaj, R. & Yu, H. The spindle checkpoint, aneuploidy, and cancer. Oncogene 23, 2016–2027 (2004).
Musacchio, A. & Salmon, E. D. The spindle-assembly checkpoint in space and time. Nat. Rev. Mol. Cell Biol. 8, 379–393 (2007).
Ciliberto, A. & Shah, J. V. A quantitative systems view of the spindle assembly checkpoint. EMBO J. 28, 2162–2173 (2009).
Vanoosthuyse, V. & Hardwick, K. G. A novel protein phosphatase 1-dependent spindle checkpoint silencing mechanism. Curr. Biol. 19, 1176–1181 (2009).
Xia, G. et al. Conformation-specific binding of p31(comet) antagonizes the function of Mad2 in the spindle checkpoint. EMBO J. 23, 3133–3143 (2004).
Reddy, S. K., Rape, M., Margansky, W. A. & Kirschner, M. W. Ubiquitination by the anaphase-promoting complex drives spindle checkpoint inactivation. Nature 446, 921–925 (2007).
Kops, G. J., Weaver, B. A. & Cleveland, D. W. On the road to cancer: aneuploidy and the mitotic checkpoint. Nat. Rev. Cancer 5, 773–785 (2005).
Schvartzman, J. M., Sotillo, R. & Benezra, R. Mitotic chromosomal instability and cancer: mouse modelling of the human disease. Nat. Rev. Cancer 10, 102–115 (2010).
Weaver, B. A. & Cleveland, D. W. The aneuploidy paradox in cell growth and tumorigenesis. Cancer Cell 14, 431–433 (2008).
Zhang, P. J. et al. CUE domain containing 2 regulates degradation of progesterone receptor by ubiquitin-proteasome. EMBO J. 26, 1831–1842 (2007).
Li, H. Y. et al. Deactivation of the kinase IKK by CUEDC2 through recruitment of the phosphatase PP1. Nat. Immunol. 9, 533–541 (2008).
Pan, J. et al. TLS-ERG leukemia fusion protein deregulates cyclin-dependent kinase 1 and blocks terminal differentiation of myeloid progenitor cells. Mol. Cancer Res. 6, 862–872 (2008).
Krasinska, L., Cot, E. & Fisher, D. Selective chemical inhibition as a tool to study Cdk1 and Cdk2 functions in the cell cycle. Cell Cycle 7, 1702–1708 (2008).
Vassilev, L. T. et al. Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc. Natl Acad. Sci. USA 103, 10660–10665 (2006).
Nicklas, R. B., Ward, S. C. & Gorbsky, G. J. Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint. J. Cell Biol. 130, 929–939 (1995).
Fang, L., Seki, A. & Fang, G. SKAP associates with kinetochores and promotes the metaphase-to-anaphase transition. Cell Cycle 8, 2819–2827 (2009).
Taylor, S. S., Ha, E. & McKeon, F. The human homologue of Bub3 is required for kinetochore localization of Bub1 and a Mad3/Bub1-related protein kinase. J. Cell Biol. 142, 1–11 (1998).
Geley, S. et al. Anaphase-promoting complex/cyclosome-dependent proteolysis of human cyclin A starts at the beginning of mitosis and is not subject to the spindle assembly checkpoint. J. Cell Biol. 153, 137–148 (2001).
den Elzen, N. & Pines, J. Cyclin A is destroyed in prometaphase and can delay chromosome alignment and anaphase. J. Cell Biol. 153, 121–136 (2001).
Fang, G., Yu, H. & Kirschner, M. W. The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation. Genes Dev. 12, 1871–1883 (1998).
Holland, A. J. & Cleveland, D. W. Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis. Nat. Rev. Mol. Cell Biol. 10, 478–487 (2009).
Ricke, R. M., van Ree, J. H. & van Deursen, J. M. Whole chromosome instability and cancer: a complex relationship. Trends Genet. 24, 457–466 (2008).
Zachariae, W. & Nasmyth, K. Whose end is destruction: cell division and the anaphase-promoting complex. Genes Dev. 13, 2039–2058 (1999).
Kraft, C. et al. Mitotic regulation of the human anaphase-promoting complex by phosphorylation. EMBO J. 22, 6598–6609 (2003).
Stegmeier, F. et al. Anaphase initiation is regulated by antagonistic ubiquitination and deubiquitination activities. Nature 446, 876–881 (2007).
Kang, R. S. et al. Solution structure of a CUE–ubiquitin complex reveals a conserved mode of ubiquitin binding. Cell 113, 621–630 (2003).
Shih, S. C. et al. A ubiquitin-binding motif required for intramolecular monoubiquitylation, the CUE domain. EMBO J. 22, 1273–1281 (2003).
Duesberg, P. & Li, R. Multistep carcinogenesis: a chain reaction of aneuploidizations. Cell Cycle 2, 202–210 (2003).
Rape, M. & Kirschner, M. W. Autonomous regulation of the anaphase-promoting complex couples mitosis to S-phase entry. Nature 432, 588–595 (2004).
Meraldi, P., Draviam, V. M. & Sorger, P. K. Timing and checkpoints in the regulation of mitotic progression. Dev. Cell 7, 45–60 (2004).
van Ree, J. H., Jeganathan, K. B., Malureanu, L. & van Deursen, J. M. Overexpression of the E2 ubiquitin-conjugating enzyme UbcH10 causeschromosome missegregation and tumor formation. J. Cell Biol. 188, 83–100 (2010).
Jin, B. F. et al. Proteomic analysis of ubiquitin–proteasome effects: insight into the function of eukaryotic initiation factor 5A. Oncogene 22, 4819–4830 (2003).
Song, M. S. et al. The tumour suppressor RASSF1A regulates mitosis by inhibiting the APC-Cdc20 complex. Nat. Cell Biol. 6, 129–137 (2004).
Meraldi, P. & Sorger, P. K. A dual role for Bub1 in the spindle checkpoint and chromosome congression. EMBO J. 24, 1621–1633 (2005).
Skoufias, D. A., Andreassen, P. R., Lacroix, F. B., Wilson, L. & Margolis, R. L. Mammalian mad2 and bub1/bubR1 recognize distinct spindle-attachment and kinetochore-tension checkpoints. Proc. Natl Acad. Sci. USA 98, 4492–4497 (2001).
Man, J. H. et al. Gankyrin plays an essential role in Ras-induced tumorigenesis through regulation of the RhoA/ROCK pathway in mammalian cells. J. Clin. Invest. 120, 2829–2841 (2010).
Hauf, S. et al. The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint. J. Cell Biol. 161, 281–294 (2003).
Acknowledgements
We thank H. T. Yu and Y. X. Zheng for discussions, M. W. Kirschner, T. Wu and G. W. Fang for technical help, Z. G. Liu for reading, S. Doxsey for HeLa cells stably expressing GFP–H2B and hTERT-RPE1 cells, E. D. Salmon for GFP–Cdc20 expression plasmid, Y. Liu for pFlag-Cdk1-AF (constitutively active) and pFlag-Cdk1-DN (kinase dead) plasmids, J. Pines for pVenus-N1 Cyclin B1 plasmid, M. W. Kirschner for the plasmids coding securin and geminin, P. K. Jackson for the plasmid pCS2-Cyclin B1, R. Benezra for the plasmid pFlag-CMV2-Mad2, H. T. Yu for pCS2-securin and pCS2-securin (ΔDB) plasmids and H. M. Wang for capturing tissue array images. This work was supported by the National Natural Science Foundation of China (no 91029733; no 30830097; no 30871234; no 30872348; no 81025010; no 30900754), the National High Technology Research and Development Program of China (2009AA02Z103 and 2009ZX09503-001) and the National Basic Research Program of China (2010CB911900 and 2010CB529904).
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X-M.Z. and H-Y.L. supervised the project; Y-F.G., Teng L. and Yan C. designed and carried out most of the experiments; Y-F.G. and Y-B.W. contributed to chromosome spread analysis; W-H.L. and K.H. analysed the phosphorylation modification with mass spectrometry; W-N.Z. and R.M. carried out immunohistochemistry analysis; Y-B.W., Tao L. and C.Z. contributed to the preparation of complementary DNA vector constructs; M.Y., Yuan C. and R.M. prepared the CUEDC2 antibody; W-L.G., B.L. and L.C. developed stable cell lines; J-H.M., Q.X. and X.P. carried out the statistics; T.Z., A-L.L., X-M.Z. and H-Y.L. analysed the data; Y-F.G., Teng L., Yan C., H-Y.L. and X-M.Z. wrote the paper.
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Gao, YF., Li, T., Chang, Y. et al. Cdk1-phosphorylated CUEDC2 promotes spindle checkpoint inactivation and chromosomal instability. Nat Cell Biol 13, 924–933 (2011). https://doi.org/10.1038/ncb2287
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DOI: https://doi.org/10.1038/ncb2287
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