Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages

  1. Weng Man Chong
  2. Won-Jing Wang
  3. Chien-Hui Lo
  4. Tzu-Yuan Chiu
  5. Ting-Jui Chang
  6. You-Pi Liu
  7. Barbara Tanos
  8. Gregory Mazo
  9. Meng-Fu Bryan Tsou
  10. Wann-Neng Jane
  11. T Tony Yang  Is a corresponding author
  12. Jung-Chi Liao  Is a corresponding author
  1. Academia Sinica, Taiwan, Republic of China
  2. National Yang-Ming University, Taiwan, Republic of China
  3. Institute of Cancer Research, United Kingdom
  4. Memorial Sloan Kettering Cancer Center, United States
  5. National Taiwan University, Taiwan, Republic of China

Abstract

Subdistal appendages (sDAPs) are centriolar elements observed proximal to the distal appendages (DAPs) in vertebrates. Despite their obvious presence, structural and functional understanding of sDAPs remains elusive. Here, by combining super-resolved localization analysis and CRISPR-Cas9 genetic perturbation, we find that, although DAPs and sDAPs are primarily responsible for distinct functions in ciliogenesis and microtubule anchoring respectively, the presence of one element actually affects the positioning of the other. Specifically, we find dual layers of both ODF2 and CEP89, where their localizations are differentially regulated by DAP and sDAP integrity. DAP depletion relaxes longitudinal occupancy of sDAP protein ninein to cover the DAP region, implying a role of DAPs in sDAP positioning. Removing sDAPs alter the distal border of centrosomal γ-tubulins, illustrating a new role of sDAPs. Together, our results provide an architectural framework of sDAPs to shed light on functional understanding, surprisingly revealing the coupling between DAPs and sDAPs.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Weng Man Chong

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  2. Won-Jing Wang

    Institute of Biochemistry and Molecular Biology, College of Life Sciences, National Yang-Ming University, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9733-0839
  3. Chien-Hui Lo

    Institute of Biochemistry and Molecular Biology, College of Life Sciences, National Yang-Ming University, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  4. Tzu-Yuan Chiu

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  5. Ting-Jui Chang

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  6. You-Pi Liu

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  7. Barbara Tanos

    Cancer Therapeutics, Institute of Cancer Research, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Gregory Mazo

    Dermatology Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Meng-Fu Bryan Tsou

    Cell Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2159-8836
  10. Wann-Neng Jane

    Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan, Republic of China
    Competing interests
    The authors declare that no competing interests exist.
  11. T Tony Yang

    Department of Electrical Engineering, National Taiwan University, Taipei, Taiwan, Republic of China
    For correspondence
    tonyyang@ntu.edu.tw
    Competing interests
    The authors declare that no competing interests exist.
  12. Jung-Chi Liao

    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan, Republic of China
    For correspondence
    jcliao@iams.sinica.edu.tw
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4323-6318

Funding

Ministry of Science and Technology, Taiwan (107-2112-M-001-037)

  • Weng Man Chong
  • Tzu-Yuan Chiu
  • Ting-Jui Chang
  • You-Pi Liu
  • T Tony Yang
  • Jung-Chi Liao

Ministry of Science and Technology, Taiwan (107-2313-B-001-009)

  • Weng Man Chong
  • Tzu-Yuan Chiu
  • Ting-Jui Chang
  • You-Pi Liu
  • T Tony Yang
  • Jung-Chi Liao

Academia Sinica (2317-1040300)

  • Weng Man Chong
  • Tzu-Yuan Chiu
  • Ting-Jui Chang
  • You-Pi Liu
  • T Tony Yang
  • Jung-Chi Liao

Ministry of Science and Technology, Taiwan (108-2313-B-010-001)

  • Won-Jing Wang

Ministry of Science and Technology, Taiwan (108-2628-B-010-007)

  • Won-Jing Wang

Ministry of Science and Technology, Taiwan (108-2638-B-010-001 -MY2)

  • Won-Jing Wang

National Institutes of Health (GM088253)

  • Meng-Fu Bryan Tsou

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

Reviewing Editor

  1. Anna Akhmanova, Utrecht University, Netherlands

Publication history

  1. Received: November 13, 2019
  2. Accepted: April 2, 2020
  3. Accepted Manuscript published: April 3, 2020 (version 1)
  4. Version of Record published: April 21, 2020 (version 2)

Copyright

© 2020, Chong 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.

Metrics

  • 3,037
    Page views
  • 543
    Downloads
  • 31
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Weng Man Chong
  2. Won-Jing Wang
  3. Chien-Hui Lo
  4. Tzu-Yuan Chiu
  5. Ting-Jui Chang
  6. You-Pi Liu
  7. Barbara Tanos
  8. Gregory Mazo
  9. Meng-Fu Bryan Tsou
  10. Wann-Neng Jane
  11. T Tony Yang
  12. Jung-Chi Liao
(2020)
Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages
eLife 9:e53580.
https://doi.org/10.7554/eLife.53580

Further reading

    1. Cell Biology
    Desiree Schatton et al.
    Research Article

    Proliferating cells undergo metabolic changes in synchrony with cell cycle progression and cell division. Mitochondria provide fuel, metabolites, and ATP during different phases of the cell cycle, however it is not completely understood how mitochondrial function and the cell cycle are coordinated. CLUH is a post-transcriptional regulator of mRNAs encoding mitochondrial proteins involved in oxidative phosphorylation and several metabolic pathways. Here, we show a role of CLUH in regulating the expression of astrin, which is involved in metaphase to anaphase progression, centrosome integrity, and mTORC1 inhibition. We find that CLUH binds both the SPAG5 mRNA and its product astrin, and controls the synthesis and the stability of the full-length astrin-1 isoform. We show that CLUH interacts with astrin-1 specifically during interphase. Astrin-depleted cells show mTORC1 hyperactivation and enhanced anabolism. On the other hand, cells lacking CLUH show decreased astrin levels and increased mTORC1 signaling, but cannot sustain anaplerotic and anabolic pathways. In absence of CLUH, cells fail to grow during G1, and progress faster through the cell cycle, indicating dysregulated matching of growth, metabolism and cell cycling. Our data reveal a role of CLUH in coupling growth signaling pathways and mitochondrial metabolism with cell cycle progression.

    1. Cell Biology
    Dillon Jevon et al.
    Research Article

    A developing understanding suggests that spatial compartmentalisation in pancreatic β cells is critical in controlling insulin secretion. To investigate the mechanisms, we have developed live-cell sub-cellular imaging methods using the mouse organotypic pancreatic slice. We demonstrate that the organotypic pancreatic slice, when compared with isolated islets, preserves intact β cell structure, and enhances glucose dependent Ca2+ responses and insulin secretion. Using the slice technique, we have discovered the essential role of local activation of integrins and the downstream component, focal adhesion kinase, in regulating β cells. Integrins and focal adhesion kinase are exclusively activated at the β cell capillary interface and using in situ and in vitro models we show their activation both positions presynaptic scaffold proteins, like ELKS and liprin, and regulates glucose dependent Ca2+ responses and insulin secretion. We conclude that focal adhesion kinase orchestrates the final steps of glucose dependent insulin secretion within the restricted domain where β cells contact the islet capillaries.