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The human mitotic checkpoint protein BubR1 regulates chromosome–spindle attachments

Nature Cell Biology volume 7, pages 93–98 (2005)Cite this article

Abstract

Loss or gain of whole chromosomes, the form of chromosomal instability (CIN) most commonly associated with human cancers, is expected to arise from the failure to accurately segregate chromosomes in mitosis1. The mitotic checkpoint is one pathway that prevents segregation errors by blocking the onset of anaphase until all chromosomes make proper attachments to the spindle. Another process that prevents errors is stabilization and destabilization of connections between chromosomes and spindle microtubules. An outstanding question is how these two pathways are coordinated to ensure accurate chromosome segregation. Here we show that in human cells depleted of BubR1 — a critical component of the mitotic checkpoint that can directly regulate the onset of anaphase2,3,4 — chromosomes do not form stable attachments to spindle microtubules. Attachments in these cells are restored by inhibition of Aurora kinase, which is known to stabilize kinetochore–microtubule attachments5,6,7. Loss of BubR1 function thus perturbs regulation of attachments rather than the ability of kinetochores to bind to microtubules. Consistent with this finding, depletion of BubR1 increases phosphorylation of CENP-A, a kinetochore-specific Aurora kinase substrate. We propose that BubR1 links regulation of chromosome–spindle attachment to mitotic checkpoint signalling.

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Figure 1: Depletion of BubR1 causes severe chromosome misalignment.
Figure 2: Depletion of BubR1 results in the loss of kinetochore–microtubule attachments.
Figure 3: Inhibition of Aurora kinase activity suppresses the BubR1-depletion phenotype.
Figure 4: BubR1 depletion increases Aurora kinase-dependent CENP-A phosphorylation at kinetochores.

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References

  1. Lengauer, C., Kinzler, K. W. & Vogelstein, B. Genetic instabilities in human cancers. Nature 396, 643–649 (1998).

    Article  CAS  Google Scholar 

  2. Sudakin, V., Chan, G. K. & Yen, T. J. Checkpoint inhibition of the APC/C in HeLa cells is mediated by a complex of BUBR1, BUB3, CDC20, and MAD2. J. Cell Biol. 154, 925–936 (2001).

    Article  CAS  Google Scholar 

  3. Tang, Z., Bharadwaj, R., Li, B. & Yu, H. Mad2-independent inhibition of APCCdc20 by the mitotic checkpoint protein BubR1. Dev. Cell 1, 227–237 (2001).

    Article  CAS  Google Scholar 

  4. Fang, G. Checkpoint protein BubR1 acts synergistically with Mad2 to inhibit anaphase-promoting complex. Mol. Biol. Cell 13, 755–766 (2002).

    Article  CAS  Google Scholar 

  5. Biggins, S. & Murray, A. W. The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint. Genes Dev. 15, 3118–3129 (2001).

    Article  CAS  Google Scholar 

  6. Tanaka, T. U. et al. Evidence that the Ipl1–Sli15 (Aurora kinase-INCENP) complex promotes chromosome bi-orientation by altering kinetochore-spindle pole connections. Cell 108, 317–329 (2002).

    Article  CAS  Google Scholar 

  7. Lampson, M. A., Renduchitala, K., Khodjakov, A. & Kapoor, T. M. Correcting improper chromosome-spindle attachments during cell division. Nature Cell Biol. 6, 232–237 (2004).

    Article  CAS  Google Scholar 

  8. Cahill, D. P. et al. Mutations of mitotic checkpoint genes in human cancers. Nature 392, 300–303 (1998).

    Article  CAS  Google Scholar 

  9. Shin, H. J. et al. Dual roles of human BubR1, a mitotic checkpoint kinase, in the monitoring of chromosomal instability. Cancer Cell 4, 483–497 (2003).

    Article  CAS  Google Scholar 

  10. Shichiri, M., Yoshinaga, K., Hisatomi, H., Sugihara, K. & Hirata, Y. Genetic and epigenetic inactivation of mitotic checkpoint genes hBUB1 and hBUBR1 and their relationship to survival. Cancer Res. 62, 13–17 (2002).

    CAS  PubMed  Google Scholar 

  11. Wang, X. et al. Significance of MAD2 expression to mitotic checkpoint control in ovarian cancer cells. Cancer Res. 62, 1662–1668 (2002).

    CAS  PubMed  Google Scholar 

  12. Hanks, S. et al. Constitutional aneuploidy and cancer predisposition caused by biallelic mutations in BUB1B. Nature Genet. 36, 1159–1161 (2004).

    Article  CAS  Google Scholar 

  13. Harrington, E. A. et al. VX-680, a potent and selective small-molecule inhibitor of the Aurora kinases, suppresses tumor growth in vivo. Nature Med. 10, 262–267 (2004).

    Article  CAS  Google Scholar 

  14. Kops, G. J., Foltz, D. R. & Cleveland, D. W. Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. Proc. Natl Acad. Sci. USA 101, 8699–8704 (2004).

    Article  CAS  Google Scholar 

  15. Li, Y. & Benezra, R. Identification of a human mitotic checkpoint gene: hsMAD2. Science 274, 246–248 (1996).

    Article  CAS  Google Scholar 

  16. Chan, G. K., Jablonski, S. A., Sudakin, V., Hittle, J. C. & Yen, T. J. Human BUBR1 is a mitotic checkpoint kinase that monitors CENP-E functions at kinetochores and binds the cyclosome/APC. J. Cell Biol. 146, 941–954 (1999).

    Article  CAS  Google Scholar 

  17. Weaver, B. A. et al. Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss. J. Cell Biol. 162, 551–563 (2003).

    Article  CAS  Google Scholar 

  18. Rieder, C. L. The structure of the cold-stable kinetochore fiber in metaphase PtK1 cells. Chromosoma 84, 145–158 (1981).

    Article  CAS  Google Scholar 

  19. King, J. M., Hays, T. S. & Nicklas, R. B. Dynein is a transient kinetochore component whose binding is regulated by microtubule attachment, not tension. J. Cell Biol. 151, 739–748 (2000).

    Article  CAS  Google Scholar 

  20. Waters, J. C., Chen, R. H., Murray, A. W. & Salmon, E. D. Localization of Mad2 to kinetochores depends on microtubule attachment, not tension. J. Cell Biol. 141, 1181–1191 (1998).

    Article  CAS  Google Scholar 

  21. Ditchfield, C. et al. Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores. J. Cell Biol. 161, 267–280 (2003).

    Article  CAS  Google Scholar 

  22. 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).

    Article  CAS  Google Scholar 

  23. Mayer, T. U. et al. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286, 971–974 (1999).

    Article  CAS  Google Scholar 

  24. Andrews, P. D. et al. Aurora B regulates MCAK at the mitotic centromere. Dev. Cell 6, 253–268 (2004).

    Article  CAS  Google Scholar 

  25. Zeitlin, S. G., Shelby, R. D. & Sullivan, K. F. CENP-A is phosphorylated by Aurora B kinase and plays an unexpected role in completion of cytokinesis. J. Cell Biol. 155, 1147–1157 (2001).

    Article  CAS  Google Scholar 

  26. Kunitoku, N. et al. CENP-A phosphorylation by Aurora-A in prophase is required for enrichment of Aurora-B at inner centromeres and for kinetochore function. Dev. Cell 5, 853–864 (2003).

    Article  CAS  Google Scholar 

  27. Earnshaw, W. C. & Rothfield, N. Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma 91, 313–321 (1985).

    Article  CAS  Google Scholar 

  28. McEwen, B. F., Heagle, A. B., Cassels, G. O., Buttle, K. F. & Rieder, C. L. Kinetochore fiber maturation in PtK1 cells and its implications for the mechanisms of chromosome congression and anaphase onset. J. Cell Biol. 137, 1567–1580 (1997).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank A. North and the Rockefeller University Bioimaging facility. We thank D. Compton, W. Dai, T. Yen, C. Walczak, W. Brinkley and D. Allis for gifts of antibodies, and H. Funabiki for discussions. This work was supported by National Institutes of Health grant GM65933 (T.M.K.). M.A.L. is a Goelet fellow.

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  1. Laboratory of Chemistry and Cell Biology, Rockefeller University, New York, 10021, NY, USA

    Michael A. Lampson & Tarun M. Kapoor

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  1. Michael A. Lampson
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Correspondence to Michael A. Lampson.

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Lampson, M., Kapoor, T. The human mitotic checkpoint protein BubR1 regulates chromosome–spindle attachments. Nat Cell Biol 7, 93–98 (2005). https://doi.org/10.1038/ncb1208

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