1. Cell Biology

Dynein-Dynactin-NuMA clusters generate cortical spindle-pulling forces as a multi-arm ensemble

  1. Masako Okumura
  2. Toyoaki Natsume
  3. Masato T Kanemaki
  4. Tomomi Kiyomitsu  Is a corresponding author
  1. Nagoya University, Japan
  2. National Institute of Genetics, Japan
https://doi.org/10.7554/eLife.36559
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  1. Masako Okumura
  2. Toyoaki Natsume
  3. Masato T Kanemaki
  4. Tomomi Kiyomitsu
(2018)
Dynein-Dynactin-NuMA clusters generate cortical spindle-pulling forces as a multi-arm ensemble
eLife 7:e36559.
https://doi.org/10.7554/eLife.36559
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Abstract

To position the mitotic spindle within the cell, dynamic plus ends of astral microtubules are pulled by membrane-associated cortical force-generating machinery. However, in contrast to the chromosome-bound kinetochore structure, how the diffusion-prone cortical machinery is organized to generate large spindle-pulling forces remains poorly understood. Here, we develop a light-induced reconstitution system in human cells. We find that induced cortical targeting of NuMA, but not dynein, is sufficient for spindle pulling. This spindle-pulling activity requires dynein-dynactin recruitment by NuMA's N-terminal long arm, dynein-based astral microtubule gliding, and NuMA's direct microtubule-binding activities. Importantly, we demonstrate that cortical NuMA assembles specialized focal structures that cluster multiple force-generating modules to generate cooperative spindle-pulling forces. This clustering activity of NuMA is required for spindle positioning, but not for spindle-pole focusing. We propose that cortical Dynein-Dynactin-NuMA (DDN) clusters act as the core force-generating machinery that organizes a multi-arm ensemble reminiscent of the kinetochore.

Data availability

All data generated or analyzed during this study are included in the manuscript, figures and supplemental files. We will deposit all plasmids and cell lines used in this study to non-profit organization such as Addgene and RIKEN BioResource Research Center.

Article and author information

Author details

  1. Masako Okumura

    Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan
    Competing interests
    The authors declare that no competing interests exist.

  2. Toyoaki Natsume

    Division of Molecular Cell Engineering, National Institute of Genetics, Mishima, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3544-4491

  3. Masato T Kanemaki

    Division of Molecular Cell Engineering, National Institute of Genetics, Mishima, Japan
    Competing interests
    The authors declare that no competing interests exist.

  4. Tomomi Kiyomitsu

    Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan
    For correspondence
    kiyomitsu@bio.nagoya-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2280-4611

Funding

Japan Science and Technology Agency (JPMJPR13A3)

  • Tomomi Kiyomitsu

Human Frontier Science Program (CDA00057/2014-C)

  • Tomomi Kiyomitsu

Japan Society for the Promotion of Science (16K14721)

  • Tomomi Kiyomitsu

Uehara Memorial Foundation

  • Tomomi Kiyomitsu

Naito Foundation

  • Tomomi Kiyomitsu

Japan Society for the Promotion of Science (17H05002)

  • Tomomi Kiyomitsu

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

Reviewing Editor

  1. Andrew P Carter, MRC Laboratory of Molecular Biology, United Kingdom

Version history

  1. Received: March 10, 2018
  2. Accepted: May 26, 2018
  3. Accepted Manuscript published: May 31, 2018 (version 1)
  4. Version of Record published: July 9, 2018 (version 2)

Copyright

© 2018, Okumura 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. Masako Okumura
  2. Toyoaki Natsume
  3. Masato T Kanemaki
  4. Tomomi Kiyomitsu
(2018)
Dynein-Dynactin-NuMA clusters generate cortical spindle-pulling forces as a multi-arm ensemble
eLife 7:e36559.
https://doi.org/10.7554/eLife.36559

Share this article

https://doi.org/10.7554/eLife.36559

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