1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics
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Multivalency of NDC80 in the outer kinetochore is essential to track shortening microtubules and generate forces

  1. Vladimir A Volkov
  2. Pim J Huis in 't Veld
  3. Marileen Dogterom  Is a corresponding author
  4. Andrea Musacchio  Is a corresponding author
  1. Delft University of Technology, Netherlands
  2. Max Planck Institute of Molecular Physiology, Germany
Research Article
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Cite as: eLife 2018;7:e36764 doi: 10.7554/eLife.36764

Abstract

Presence of multiple copies of the microtubule-binding NDC80 complex is an evolutionary conserved feature of kinetochores, points of attachment of chromosomes to spindle microtubules. This may enable multivalent interactions with microtubules, with implications that remain unexplored. Using recombinant human kinetochore components, we show that while single NDC80 complexes do not track depolymerizing microtubules, reconstituted particles containing the NDC80 receptor CENP-T bound to three or more NDC80 complexes do so effectively, as expected for a kinetochore force coupler. To study multivalency systematically, we engineered modules allowing incremental addition of NDC80 complexes. The modules' residence time on microtubules increased exponentially with the number of NDC80 complexes. Modules with two or more complexes tracked depolymerizing microtubules with increasing efficiencies, and stalled and rescued microtubule depolymerization in a force-dependent manner when conjugated to cargo. Our observations indicate that NDC80, rather than through biased diffusion, tracks depolymerizing microtubules by harnessing force generated during microtubule disassembly.

Article and author information

Author details

  1. Vladimir A Volkov

    Department of Bionanoscience, Delft University of Technology, Delft, Netherlands
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0002-5407-3366
  2. Pim J Huis in 't Veld

    Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0003-0234-6390
  3. Marileen Dogterom

    Department of Bionanoscience, Delft University of Technology, Delft, Netherlands
    For correspondence
    m.dogterom@tudelft.nl
    Competing interests
    No competing interests declared.
  4. 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, Senior editor, eLife.
    ORCID icon 0000-0003-2362-8784

Funding

H2020 European Research Council (669686)

  • Andrea Musacchio

H2020 European Research Council (609822)

  • Marileen Dogterom

European Molecular Biology Organization (7203)

  • Pim J Huis in 't Veld

Deutsche Forschungsgemeinschaft (CRC1093)

  • Andrea Musacchio

Max-Planck-Gesellschaft (Open-access funding)

  • Andrea Musacchio

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

Reviewing Editor

  1. Jessica K Tyler, Reviewing Editor, Weill Cornell Medicine, United States

Publication history

  1. Received: March 17, 2018
  2. Accepted: March 31, 2018
  3. Accepted Manuscript published: April 9, 2018 (version 1)

Copyright

© 2018, Volkov 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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