Aurora B phosphorylates centromeric MCAK and regulates its localization and microtubule depolymerization activity

Weijie Lan, Xin Zhang, Susan L. Kline-Smith, Sara E. Rosasco, Gregory A. Barrett-Wilt, Jeffrey Shabanowitz, Donald F. Hunt, Claire E. Walczak, P. Todd Stukenberg

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353 Scopus citations

Abstract

Background: Sister kinetochores must bind microtubules in a bipolar fashion to equally segregate chromosomes during mitosis. The molecular mechanisms underlying this process remain unclear. Aurora B likely promotes chromosome biorientation by regulating kinetochore-microtubule attachments. MCAK (mitotic centromere-associated kinesin) is a Kin I kinesin that can depolymerize microtubules. These two proteins both localize to mitotic centromeres and have overlapping mitotic functions, including regulation of microtubule dynamics, proper chromosome congression, and correction of improper kinetochore-microtubule attachments. Results: We show that Aurora B phosphorylates and regulates MCAK both in vitro and in vivo. Specifically, we mapped six Aurora B phosphorylation sites on MCAK in both the centromere-targeting domain and the neck region. Aurora B activity was required to localize MCAK to centromeres, but not to spindle poles. Aurora B phosphorylation of serine 196 in the neck region of MCAK inhibited its microtubule depolymerization activity. We found that this key site was phosphorylated at centromeres and anaphase spindle midzones in vivo. However, within the inner centromere there were pockets of both phosphorylated and unphosphorylated MCAK protein, suggesting that phosphate turnover is crucial in the regulation of MCAK activity. Addition of α-p-S196 antibodies to Xenopus egg extracts or injection of α-p-S196 antibodies into cells caused defects in chromosome positioning and/or segregation. Conclusions: We have established a direct link between the microtubule depolymerase MCAK and Aurora B kinase. Our data suggest that Aurora B both positively and negatively regulates MCAK during mitosis. We propose that Aurora B biorients chromosomes by directing MCAK to depolymerize incorrectly oriented kinetochore microtubules.

Original languageEnglish (US)
Pages (from-to)273-286
Number of pages14
JournalCurrent Biology
Volume14
Issue number4
DOIs
StatePublished - Feb 17 2004

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Lan, W., Zhang, X., Kline-Smith, S. L., Rosasco, S. E., Barrett-Wilt, G. A., Shabanowitz, J., Hunt, D. F., Walczak, C. E., & Stukenberg, P. T. (2004). Aurora B phosphorylates centromeric MCAK and regulates its localization and microtubule depolymerization activity. Current Biology, 14(4), 273-286. https://doi.org/10.1016/S0960-9822(04)00064-8