Systematic studies of all PIH proteins in zebrafish reveal their distinct roles in axonemal dynein assembly

  1. Hiroshi Yamaguchi
  2. Toshiyuki Oda
  3. Masahide Kikkawa  Is a corresponding author
  4. Hiroyuki Takeda  Is a corresponding author
  1. The University of Tokyo, Japan
  2. University of Yamanashi, Japan

Abstract

Construction of motile cilia/flagella requires cytoplasmic preassembly of axonemal dyneins before transport into cilia. Axonemal dyneins have various subtypes, but the roles of each dynein subtype and their assembly processes remain elusive in vertebrates. The PIH protein family, consisting of four members, has been implicated in the assembly of different dynein subtypes, although evidence for this idea is sparse. Here, we established zebrafish mutants of all four PIH-protein genes: pih1d1, pih1d2, ktu, and twister, and analyzed the structures of axonemal dyneins in mutant spermatozoa by cryo-electron tomography. Mutations caused the loss of specific dynein subtypes, which was correlated with abnormal sperm motility. We also found organ-specific compositions of dynein subtypes, which could explain the severe motility defects of mutant Kupffer's vesicle cilia. Our data demonstrate that all vertebrate PIH proteins are differently required for cilia/flagella motions and the assembly of axonemal dyneins, assigning specific dynein subtypes to each PIH protein.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. All the electron density maps derived from cryo-electron tomography are deposited in EMD under the following accession numbers: WT, EMD-6954; pih1d1-/-, EMD-6955; pih1d2-/-, EMD-6956; ktu-/-, EMD-6957; twister-/-, EMD-6958; pih1d2-/-;ktu-/- (+OAD class), EMD-6959; and pih1d2-/-;ktu-/- (-OAD class), EMD-6960.

Article and author information

Author details

  1. Hiroshi Yamaguchi

    Department of Cell Biology and Anatomy, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8722-129X
  2. Toshiyuki Oda

    Department of Anatomy and Structural Biology, Graduate School of Medical Science, University of Yamanashi, Chuo, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8090-2159
  3. Masahide Kikkawa

    Department of Cell Biology and Anatomy, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Japan
    For correspondence
    mkikkawa@m.u-tokyo.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7656-8194
  4. Hiroyuki Takeda

    Department of Biological Sciences, Graduate School of Science,, The University of Tokyo, Bunkyo-ku, Japan
    For correspondence
    htakeda@bs.s.u-tokyo.ac.jp
    Competing interests
    The authors declare that no competing interests exist.

Funding

Core Research for Evolutional Science and Technology (JPMJCR14M1)

  • Masahide Kikkawa

Japan Society for the Promotion of Science (16H02502)

  • Masahide Kikkawa

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: March 26, 2018
  2. Accepted: May 5, 2018
  3. Accepted Manuscript published: May 9, 2018 (version 1)
  4. Version of Record published: June 19, 2018 (version 2)

Copyright

© 2018, Yamaguchi 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. Hiroshi Yamaguchi
  2. Toshiyuki Oda
  3. Masahide Kikkawa
  4. Hiroyuki Takeda
(2018)
Systematic studies of all PIH proteins in zebrafish reveal their distinct roles in axonemal dynein assembly
eLife 7:e36979.
https://doi.org/10.7554/eLife.36979

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