Molecular basis of outer kinetochoreassembly on CENP-T

  1. Pim J Huis in 't Veld  Is a corresponding author
  2. Sadasivam Jeganathan
  3. Arsen Petrovic
  4. Priyanka Singh
  5. Juliane John
  6. Veronica Krenn
  7. Florian Weissmann
  8. Tanja Bange
  9. Andrea Musacchio  Is a corresponding author
  1. Max Planck Institute of Molecular Physiology, Germany
  2. Chemical Genomics Centre of the Max Planck Society, Germany
  3. Vienna Biocenter, Austria
  4. Vienna Biocenter, Germany

Abstract

Stable kinetochore-microtubule attachment is essential for cell division. It requires recruitment of outer kinetochore microtubule binders by centromere proteins C and T (CENP-C and CENP-T). To study the molecular requirements of kinetochore formation, we reconstituted the binding of the MIS12 and NDC80 outer kinetochore subcomplexes to CENP-C and CENP-T. Whereas CENP-C recruits a single MIS12:NDC80 complex, we show here that CENP-T binds one MIS12:NDC80 and two NDC80 complexes upon phosphorylation by the mitotic CDK1:Cyclin B complex at three distinct CENP-T sites. Visualization of reconstituted complexes by electron microscopy supports this model. Binding of CENP-C and CENP-T to MIS12 is competitive, and therefore CENP-C and CENP-T act in parallel to recruit two MIS12 and up to four NDC80 complexes. Our observations provide a molecular explanation for the stoichiometry of kinetochore components and its cell cycle regulation, and highlight how outer kinetochore modules bridge distances of well over 100 nm.

Article and author information

Author details

  1. Pim J Huis in 't Veld

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    For correspondence
    pim.huis@mpi-dortmund.mpg.de
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0234-6390
  2. Sadasivam Jeganathan

    Chemical Genomics Centre of the Max Planck Society, Dortmund, Germany
    Competing interests
    No competing interests declared.
  3. Arsen Petrovic

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    Competing interests
    No competing interests declared.
  4. Priyanka Singh

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    Competing interests
    No competing interests declared.
  5. Juliane John

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    Competing interests
    No competing interests declared.
  6. Veronica Krenn

    Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna Biocenter, Vienna, Austria
    Competing interests
    No competing interests declared.
  7. Florian Weissmann

    Research Institute of Molecular Pathology, Vienna Biocenter, Vienna, Germany
    Competing interests
    No competing interests declared.
  8. Tanja Bange

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    Competing interests
    No competing interests declared.
  9. Andrea Musacchio

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    For correspondence
    andrea.musacchio@mpi-dortmund.mpg.de
    Competing interests
    Andrea Musacchio, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2362-8784

Funding

European Research Council (AdG 669686 RECEPIANCE)

  • Andrea Musacchio

Deutsche Forschungsgemeinschaft (CRC1093)

  • Andrea Musacchio

European Molecular Biology Organization (ALTF 262-2009)

  • Sadasivam Jeganathan

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Jon Pines, The Gurdon Institute, United Kingdom

Version history

  1. Received: August 26, 2016
  2. Accepted: December 23, 2016
  3. Accepted Manuscript published: December 24, 2016 (version 1)
  4. Accepted Manuscript updated: December 28, 2016 (version 2)
  5. Version of Record published: January 17, 2017 (version 3)

Copyright

© 2016, Huis in 't Veld et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Pim J Huis in 't Veld
  2. Sadasivam Jeganathan
  3. Arsen Petrovic
  4. Priyanka Singh
  5. Juliane John
  6. Veronica Krenn
  7. Florian Weissmann
  8. Tanja Bange
  9. Andrea Musacchio
(2016)
Molecular basis of outer kinetochoreassembly on CENP-T
eLife 5:e21007.
https://doi.org/10.7554/eLife.21007

Share this article

https://doi.org/10.7554/eLife.21007

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