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The mitotic checkpoint kinase NEK2A regulates kinetochore microtubule attachment stability

Abstract

Loss or gain of whole chromosome, the form of chromosome instability commonly associated with cancers is thought to arise from aberrant chromosome segregation during cell division. Chromosome segregation in mitosis is orchestrated by the interaction of kinetochores with spindle microtubules. Our studies show that NEK2A is a kinetochore-associated protein kinase essential for faithful chromosome segregation. However, it was unclear how NEK2A ensures accurate chromosome segregation in mitosis. Here we show that NEK2A-mediated Hec1 (highly expressed in cancer) phosphorylation is essential for faithful kinetochore microtubule attachments in mitosis. Using phospho-specific antibody, our studies show that NEK2A phosphorylates Hec1 at Ser165 during mitosis. Although such phosphorylation is not required for assembly of Hec1 to the kinetochore, expression of non-phosphorylatable mutant Hec1S165 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic and monotelic attachments. Our in vitro reconstitution experiment demonstrated that Hec1 binds to microtubule in low affinity and phosphorylation by NEK2A, which prevents aberrant kinetochore-microtubule connections in vivo, increases the affinity of the Ndc80 complex for microtubules in vitro. Thus, our studies illustrate a novel regulatory mechanism in which NEK2A kinase operates a faithful chromosome attachment to spindle microtubule, which prevents chromosome instability during cell division.

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Abbreviations

Hec1:

highly expressed in cancer

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Acknowledgements

We would like to thank Drs W-H Lee and A Musacchio for gift reagents. This work was supported by grants from Chinese 973 project (2002CB713700, 2007CB914503), Chinese Academy of Sciences (KSCX1-YW-R65 and KSCX2-YW-H10), Chinese 863 project (2001AA215331 and), Chinese Ministry of Education (20020358051; 20050358061 and 111 project B07007 to XD), Chinese Natural Science Foundation (90508002, 30121001 to XY; 30500183 to XD; 30570850 to JZ; 30600222 to JY), National Institutes of Health (DK56292, CA132389, CA89019, and CA92080) and a GCC Breast Cancer Research Grant to XY. The facilities were supported in part by NIH/NCRR/RCM1 Grant G-12-RR03034. SER and XY are GCC eminent scholars.

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Correspondence to X Ding, C Jin or X Yao.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Du, J., Cai, X., Yao, J. et al. The mitotic checkpoint kinase NEK2A regulates kinetochore microtubule attachment stability. Oncogene 27, 4107–4114 (2008). https://doi.org/10.1038/onc.2008.34

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Keywords

  • chromosome instability
  • mitosis
  • kinetochore-microtubule attachment
  • Hec1
  • NEK2A
  • phosphorylation

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