1. Structural Biology and Molecular Biophysics

Recruitment of two dyneins to an mRNA-dependent Bicaudal D transport complex

  1. Thomas E Sladewski
  2. Neil Billington
  3. M Yusuf Ali
  4. Carol S Bookwalter
  5. Hailong Lu
  6. Elena B Krementsova
  7. Trina A Schroer
  8. Kathleen M Trybus  Is a corresponding author
  1. University of Vermont, United States
  2. National Heart, Lung and Blood Institute, National Institutes of Health, United States
  3. Johns Hopkins University, United States
https://doi.org/10.7554/eLife.36306
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  1. Thomas E Sladewski
  2. Neil Billington
  3. M Yusuf Ali
  4. Carol S Bookwalter
  5. Hailong Lu
  6. Elena B Krementsova
  7. Trina A Schroer
  8. Kathleen M Trybus
(2018)
Recruitment of two dyneins to an mRNA-dependent Bicaudal D transport complex
eLife 7:e36306.
https://doi.org/10.7554/eLife.36306
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Abstract

We investigated the role of full-length Drosophila Bicaudal D (BicD) binding partners in dynein-dynactin activation for mRNA transport on microtubules. Full-length BicD robustly activated dynein-dynactin motility only when both the mRNA binding protein Egalitarian (Egl) and K10 mRNA cargo were present, and electron microscopy showed that both Egl and mRNA were needed to disrupt a looped, auto-inhibited BicD conformation. BicD can recruit two dimeric dyneins, resulting in faster speeds and longer runs than with one dynein. Moving complexes predominantly contained two Egl molecules and one K10 mRNA. This mRNA-bound configuration makes Egl bivalent, likely enhancing its avidity for BicD and thus its ability to disrupt BicD auto-inhibition. Consistent with this idea, artificially dimerized Egl activates dynein-dynactin-BicD in the absence of mRNA. The ability of mRNA cargo to orchestrate the activation of the mRNP (messenger ribonucleotide protein) complex is an elegant way to ensure that only cargo-bound motors are motile.

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Data generated or analyzed during this study are included in the manuscript and supporting files. Source data files have been included for Figures 1, 4-10.

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Author details

  1. Thomas E Sladewski

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Neil Billington

    Laboratory of Physiology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2306-0228

  3. M Yusuf Ali

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Carol S Bookwalter

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Hailong Lu

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Elena B Krementsova

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Trina A Schroer

    Department of Biology, Johns Hopkins University, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5065-1835

  8. Kathleen M Trybus

    Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, United States
    For correspondence
    kathleen.trybus@uvm.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5583-8500

Funding

National Institutes of Health (GM078097)

  • Kathleen M Trybus

American Heart Association (12SDG11930002)

  • M Yusuf Ali

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

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

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  1. Thomas E Sladewski
  2. Neil Billington
  3. M Yusuf Ali
  4. Carol S Bookwalter
  5. Hailong Lu
  6. Elena B Krementsova
  7. Trina A Schroer
  8. Kathleen M Trybus
(2018)
Recruitment of two dyneins to an mRNA-dependent Bicaudal D transport complex
eLife 7:e36306.
https://doi.org/10.7554/eLife.36306

Share this article

https://doi.org/10.7554/eLife.36306

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Further reading

    1. Cell Biology

    RNA-directed activation of cytoplasmic dynein-1 in reconstituted transport RNPs

    Mark A McClintock, Carly I Dix ... Simon L Bullock
    Research Article Updated

    Polarised mRNA transport is a prevalent mechanism for spatial control of protein synthesis. However, the composition of transported ribonucleoprotein particles (RNPs) and the regulation of their movement are poorly understood. We have reconstituted microtubule minus end-directed transport of mRNAs using purified components. A Bicaudal-D (BicD) adaptor protein and the RNA-binding protein Egalitarian (Egl) are sufficient for long-distance mRNA transport by the dynein motor and its accessory complex dynactin, thus defining a minimal transport-competent RNP. Unexpectedly, the RNA is required for robust activation of dynein motility. We show that a cis-acting RNA localisation signal promotes the interaction of Egl with BicD, which licenses the latter protein to recruit dynein and dynactin. Our data support a model for BicD activation based on RNA-induced occupancy of two Egl-binding sites on the BicD dimer. Scaffolding of adaptor protein assemblies by cargoes is an attractive mechanism for regulating intracellular transport.