1. Cell Biology

Centriole triplet microtubules are required for stable centriole formation and inheritance in human cells

  1. Jennifer T Wang
  2. Dong Kong
  3. Christian R Hoerner
  4. Jadranka Loncarek
  5. Tim Stearns  Is a corresponding author
  1. Stanford University, United States
  2. National Cancer Institute, National Institutes of Health, United States
  3. Stanford School of Medicine, United States
https://doi.org/10.7554/eLife.29061
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  1. Jennifer T Wang
  2. Dong Kong
  3. Christian R Hoerner
  4. Jadranka Loncarek
  5. Tim Stearns
(2017)
Centriole triplet microtubules are required for stable centriole formation and inheritance in human cells
eLife 6:e29061.
https://doi.org/10.7554/eLife.29061
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Abstract

Centrioles are composed of long-lived microtubules arranged in nine triplets. However, the contribution of triplet microtubules to mammalian centriole formation and stability is unknown. Little is known of the mechanism of triplet microtubule formation, but experiments in unicellular eukaryotes indicate that delta-tubulin and epsilon-tubulin, two less-studied tubulin family members, are required. Here, we report that centrioles in delta-tubulin and epsilon-tubulin null mutant human cells lack triplet microtubules and fail to undergo centriole maturation. These aberrant centrioles are formed de novo each cell cycle, but are unstable and do not persist to the next cell cycle, leading to a futile cycle of centriole formation and­­­ disintegration. Disintegration can be suppressed by paclitaxel treatment. Delta-tubulin and epsilon-tubulin physically interact, indicating that these tubulins act together to maintain triplet microtubules and that these are necessary for inheritance of centrioles from one cell cycle to the next

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Author details

  1. Jennifer T Wang

    Department of Biology, Stanford University, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8506-5182

  2. Dong Kong

    Laboratory of Protein Dynamics and Signaling, Center for Cancer Research - Frederick, National Cancer Institute, National Institutes of Health, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Christian R Hoerner

    Division of Oncology, Department of Medicine, Stanford School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Jadranka Loncarek

    Laboratory of Protein Dynamics and Signaling, Center for Cancer Research - Frederick, National Cancer Institute, National Institutes of Health, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Tim Stearns

    Department of Biology, Stanford University, Stanford, United States
    For correspondence
    stearns@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0671-6582

Funding

National Institute of General Medical Sciences (5 F32 GM117678)

  • Jennifer T Wang

National Cancer Institute (Intramural Program)

  • Dong Kong
  • Jadranka Loncarek

National Institute of General Medical Sciences (R01GM052022)

  • Tim Stearns

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. Jennifer T Wang
  2. Dong Kong
  3. Christian R Hoerner
  4. Jadranka Loncarek
  5. Tim Stearns
(2017)
Centriole triplet microtubules are required for stable centriole formation and inheritance in human cells
eLife 6:e29061.
https://doi.org/10.7554/eLife.29061

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https://doi.org/10.7554/eLife.29061

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    1. Cell Biology

    A delta-tubulin/epsilon-tubulin/Ted protein complex is required for centriole architecture

    Rachel Pudlowski, Lingyi Xu ... Jennifer T Wang
    Research Advance

    Centrioles have a unique, conserved architecture formed by three linked, ‘triplet’, microtubules arranged in ninefold symmetry. The mechanisms by which these triplet microtubules are formed remain unclear but likely involve the noncanonical tubulins delta-tubulin and epsilon-tubulin. Previously, we found that human cells lacking delta-tubulin or epsilon-tubulin form abnormal centrioles, characterized by an absence of triplet microtubules, lack of central core protein POC5, and a futile cycle of centriole formation and disintegration (Wang et al., 2017). Here, we show that human cells lacking either TEDC1 or TEDC2 have similar abnormalities. Using ultrastructure expansion microscopy, we observed that mutant centrioles elongate to the same length as control centrioles in G2 phase and fail to recruit central core scaffold proteins. Remarkably, mutant centrioles also have an expanded proximal region. During mitosis, these mutant centrioles further elongate before fragmenting and disintegrating. All four proteins physically interact and TEDC1 and TEDC2 can form a subcomplex in the absence of the tubulins, supporting an AlphaFold Multimer model of the tetramer. TEDC1 and TEDC2 localize to centrosomes and are mutually dependent on each other and on delta-tubulin and epsilon-tubulin for localization. Our results demonstrate that delta-tubulin, epsilon-tubulin, TEDC1, and TEDC2 function together to promote robust centriole architecture, laying the foundation for future studies on the mechanisms underlying the assembly of triplet microtubules and their interactions with centriole structure.