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Myosin-Va is required for preciliary vesicle transportation to the mother centriole during ciliogenesis

Nature Cell Biologyvolume 20pages175185 (2018) | Download Citation

Abstract

Primary cilia play essential roles in signal transduction and development. The docking of preciliary vesicles at the distal appendages of a mother centriole is an initial/critical step of ciliogenesis, but the mechanisms are unclear. Here, we demonstrate that myosin-Va mediates the transportation of preciliary vesicles to the mother centriole and reveal the underlying mechanism. We also show that the myosin-Va-mediated transportation of preciliary vesicles is the earliest event that defines the onset of ciliogenesis. Depletion of myosin-Va significantly inhibits the attachment of preciliary vesicles to the distal appendages of the mother centriole and decreases cilia assembly. Myosin-Va functions upstream of EHD1- and Rab11-mediated ciliary vesicle formation. Importantly, dynein mediates myosin-Va-associated preciliary vesicle transportation to the pericentrosomal region along microtubules, while myosin-Va mediates preciliary vesicle transportation from the pericentrosomal region to the distal appendages of the mother centriole via the Arp2/3-associated branched actin network.

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Acknowledgements

The authors acknowledge support from the sequencing core facility (IBMS), the confocal imaging core facilities (IBMS, IMB, NPAS) and the EM core facilities (IMB, ICOB) of Academia Sinica. This work was supported by grants from the Ministry of Science and Technology, Taiwan (MOST 105-2321-B001-016) and the Academia Sinica Investigator Award.

Author information

Affiliations

  1. Taiwan International Graduate Program in Interdisciplinary Neuroscience, National Yang-Ming University and Academia Sinica, Taipei, Taiwan

    • Chien-Ting Wu
    •  & Tang K. Tang
  2. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    • Chien-Ting Wu
    • , Hsin-Yi Chen
    •  & Tang K. Tang

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Contributions

C.-T.W., a PhD student at the Taiwan International Graduate Program in Interdisciplinary Neuroscience, National Yang-Ming University and Academia Sinica, performed most of the experiments, designed the study, interpreted data and wrote the initial draft of the manuscript. H.-Y.C. performed experiments. T.K.T. conceived and designed the study, interpreted the data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Tang K. Tang.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–8, Supplementary legends.

  2. Life Sciences Reporting Summary

  3. Supplementary Table 1

    List of antibodies and dilutions used in this study.

  4. Supplementary Table 2

    List of siRNA sequences used in this study.

Videos

  1. Supplementary Video 1

    Related to Supplementary Fig. 2b. RPE1-based inducible cells expressing GFP-Myo-Va-GTD and mCherry-Arl13b were treated with DOX for 24 h and then serum-starved. Live cell images were taken using a LSM780 Carl Zeiss confocal system under controlled CO2 (5%) and temperature (37 °C).

  2. Supplementary Video 2

    Related to Supplementary Fig. 2d. NIH3T3-based inducible cells expressing GFP-Myo-Va-GTD and mCherry-Arl13b were treated with DOX for 24 h and then serum-starved. Live cell images were taken using a LSM780 Carl Zeiss confocal system under controlled CO2 (5%) and temperature (37 °C).

  3. Supplementary Video 3

    Related to Fig. 2f. IMCD3-based inducible cells expressing GFP-Myo-Va-GTD and mCherry-Arl13b were treated with DOX for 24 h and then serum-starved. Live cell images were taken using a LSM780 Carl Zeiss confocal system under controlled CO2 (5%) and temperature (37 °C).

  4. Supplementary Video 4

    Related to Supplementary Fig. 1e. RPE1-based inducible cells expressing GFP-EHD1 and mCherry-Myo-Va-GTD were treated with DOX for 24 h and then serum-starved. Live cell images were taken using a LSM780 Carl Zeiss confocal system under controlled CO2 (5%) and temperature (37 °C).

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DOI

https://doi.org/10.1038/s41556-017-0018-7