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Optogenetic dissection of mitotic spindle positioning in vivo

  1. Lars-Eric Fielmich
  2. Ruben Schmidt
  3. Daniel J Dickinson
  4. Bob Goldstein
  5. Anna Akhmanova
  6. Sander Van den Heuvel  Is a corresponding author
  1. Utrecht University, Netherlands
  2. University of North Carolina at Chapel Hill, United States
Research Article
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Cite this article as: eLife 2018;7:e38198 doi: 10.7554/eLife.38198

Abstract

The position of the mitotic spindle determines the plane of cell cleavage, and thereby daughter cell location, size, and content. Spindle positioning is driven by dynein-mediated pulling forces exerted on astral microtubules, which requires an evolutionarily conserved complex of Gα-GDP, GPR-1/2Pins/LGN, and LIN-5Mud/NuMA proteins. To examine individual functions of the complex components, we developed a genetic strategy for light-controlled localization of endogenous proteins in C. elegans embryos. By replacing Gα and GPR-1/2 with a light-inducible membrane anchor, we demonstrate that Gα-GDP, Gα-GTP, and GPR-1/2 are not required for pulling-force generation. In the absence of Gα and GPR-1/2, cortical recruitment of LIN-5, but not dynein itself, induced high pulling forces. The light-controlled localization of LIN-5 overruled normal cell-cycle and polarity regulation and provided experimental control over the spindle and cell-cleavage plane. Our results define Gα∙GDP-GPR-1/2 Pins/LGN as a regulatable membrane anchor, and LIN-5Mud/NuMA as a potent activator of dynein-dependent spindle-positioning forces.

Article and author information

Author details

  1. Lars-Eric Fielmich

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0247-1298
  2. Ruben Schmidt

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9187-5424
  3. Daniel J Dickinson

    Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  4. Bob Goldstein

    Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    No competing interests declared.
  5. Anna Akhmanova

    Department of Biology, Utrecht University, Utrecht, Netherlands
    Competing interests
    Anna Akhmanova, Deputy editor of eLife Magazine.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9048-8614
  6. Sander Van den Heuvel

    Department of Biology, Utrecht University, Utrecht, Netherlands
    For correspondence
    s.j.l.vandenheuvel@uu.nl
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9015-7463

Funding

Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO TOP/ECHO grant 711.015.001)

  • Lars-Eric Fielmich
  • Sander Van den Heuvel

National Institutes of Health (NIH R01 GM083071)

  • Bob Goldstein

Helen Hay Whitney Foundation

  • Daniel J Dickinson

European Research Council (Synergy grant 609822)

  • Ruben Schmidt
  • Anna Akhmanova

National Institutes of Health (NIH K99 GM115964)

  • Daniel J Dickinson

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

Publication history

  1. Received: May 9, 2018
  2. Accepted: August 14, 2018
  3. Accepted Manuscript published: August 15, 2018 (version 1)
  4. Version of Record published: November 2, 2018 (version 2)

Copyright

© 2018, Fielmich 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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