Centriolar satellites expedite mother centriole remodeling to promote ciliogenesis

Abstract

Centrosomes are orbited by centriolar satellites, dynamic multiprotein assemblies nucleated by Pericentriolar Material 1 (PCM1). To study the requirement for centriolar satellites, we generated mice lacking PCM1, a crucial component of satellites. Pcm1-/- mice display partially penetrant perinatal lethality with survivors exhibiting hydrocephalus, oligospermia and cerebellar hypoplasia, and variably expressive phenotypes such as hydronephrosis. As many of these phenotypes have been observed in human ciliopathies and satellites are implicated in cilia biology, we investigated whether cilia were affected. PCM1 was dispensable for ciliogenesis in many cell types, whereas Pcm1-/- multiciliated ependymal cells and human PCM1-/- retinal pigmented epithelial 1 (RPE1) cells showed reduced ciliogenesis. PCM1-/- RPE1 cells displayed reduced docking of the mother centriole to the ciliary vesicle and removal of CP110 and CEP97 from the distal mother centriole, indicating compromised early ciliogenesis. Similarly, Pcm1-/- ependymal cells exhibited reduced removal of CP110 from basal bodies in vivo. We propose that PCM1 and centriolar satellites facilitate efficient trafficking of proteins to and from centrioles, including the departure of CP110 and CEP97 to initiate ciliogenesis, and that the threshold to trigger ciliogenesis differs between cell types.

Data availability

Proteomics data files are be uploaded ProteomeXchange (Identifier: PXD031920), with the accession number is available with the paper.All analysis tools have been made available on GitHub (https://github.com/IGC-Advanced-Imaging-Resource/Hall2022_Paper), as described in Materials and Methods.

The following data sets were generated

Article and author information

Author details

  1. Emma A Hall

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  2. Dhivya Kumar

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3737-014X
  3. Suzanna L Prosser

    Lunenfeld-Tanenbaum Research Institute, Toronto, Canada
    Competing interests
    No competing interests declared.
  4. Patricia L Yeyati

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  5. Vicente Herranz-Pérez

    Cavanilles Institute of Biodiversity and Evolutionary Biology, University of Valencia, Valencia, Spain
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1969-1214
  6. Jose Manuel García-Verdugo

    Cavanilles Institute of Biodiversity and Evolutionary Biology, University of Valencia, Valencia, Spain
    Competing interests
    No competing interests declared.
  7. Lorraine Rose

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  8. Lisa McKie

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  9. Daniel O Dodd

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  10. Peter A Tennant

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  11. Roly Megaw

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5605-4540
  12. Laura C Murphy

    Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  13. Marisa F Ferreira

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8123-4612
  14. Graeme Grimes

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  15. Lucy Williams

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
  16. Tooba Quidwai

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5248-9010
  17. Laurence Pelletier

    Lunenfeld-Tanenbaum Research Institute, Toronto, Canada
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1171-4618
  18. Jeremy F Reiter

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    For correspondence
    Jeremy.Reiter@ucsf.edu
    Competing interests
    Jeremy F Reiter, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6512-320X
  19. Pleasantine Mill

    MRC Human Genetics Unit, University of Edinburgh, Edinburgh, United Kingdom
    For correspondence
    pleasantine.mill@ed.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5218-134X

Funding

Medical Research Council (MR_UU_1201018/26)

  • Emma A Hall
  • Dhivya Kumar
  • Patricia L Yeyati
  • Lorraine Rose
  • Lisa McKie
  • Daniel O Dodd
  • Peter A Tennant
  • Roly Megaw
  • Laura C Murphy
  • Marisa F Ferreira
  • Graeme Grimes
  • Lucy Williams
  • Tooba Quidwai
  • Pleasantine Mill

Sandler Foundation

  • Dhivya Kumar

Krembil Foundation

  • Suzanna L Prosser
  • Laurence Pelletier

European Commission (866355)

  • Emma A Hall
  • Daniel O Dodd
  • Pleasantine Mill

Canadian Institutes of Health Research (167279)

  • Suzanna L Prosser
  • Laurence Pelletier

European Commission (702601)

  • Suzanna L Prosser

National Institutes of Health (R01GM095941)

  • Dhivya Kumar
  • Vicente Herranz-Pérez
  • Jose Manuel García-Verdugo
  • Jeremy F Reiter

National Institutes of Health (R01AR054396)

  • Dhivya Kumar
  • Vicente Herranz-Pérez
  • Jose Manuel García-Verdugo
  • Jeremy F Reiter

National Institutes of Health (RO1HD089918)

  • Dhivya Kumar
  • Vicente Herranz-Pérez
  • Jose Manuel García-Verdugo
  • Jeremy F Reiter

National Institutes of Health (5K99GM140175)

  • Dhivya Kumar

Jane Coffin Childs Memorial Fund for Medical Research

  • Dhivya Kumar

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

Ethics

Animal experimentation: Animals were maintained in SPF environment and studies carried out in accordance with the guidance issued by the Medical Research Council in "Responsibility in the Use of Animals in Medical Research" (July 1993) and licensed by the Home Office under the Animals (Scientific Procedures) Act 1986 under project license number P18921CDE in facilities at the University of Edinburgh (PEL 60/6025).

Copyright

© 2023, Hall 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

  • 3,199
    views
  • 554
    downloads
  • 29
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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. Emma A Hall
  2. Dhivya Kumar
  3. Suzanna L Prosser
  4. Patricia L Yeyati
  5. Vicente Herranz-Pérez
  6. Jose Manuel García-Verdugo
  7. Lorraine Rose
  8. Lisa McKie
  9. Daniel O Dodd
  10. Peter A Tennant
  11. Roly Megaw
  12. Laura C Murphy
  13. Marisa F Ferreira
  14. Graeme Grimes
  15. Lucy Williams
  16. Tooba Quidwai
  17. Laurence Pelletier
  18. Jeremy F Reiter
  19. Pleasantine Mill
(2023)
Centriolar satellites expedite mother centriole remodeling to promote ciliogenesis
eLife 12:e79299.
https://doi.org/10.7554/eLife.79299

Share this article

https://doi.org/10.7554/eLife.79299

Further reading

    1. Cell Biology
    2. Evolutionary Biology
    Paul Richard J Yulo, Nicolas Desprat ... Heather L Hendrickson
    Research Article

    Maintenance of rod-shape in bacterial cells depends on the actin-like protein MreB. Deletion of mreB from Pseudomonas fluorescens SBW25 results in viable spherical cells of variable volume and reduced fitness. Using a combination of time-resolved microscopy and biochemical assay of peptidoglycan synthesis, we show that reduced fitness is a consequence of perturbed cell size homeostasis that arises primarily from differential growth of daughter cells. A 1000-generation selection experiment resulted in rapid restoration of fitness with derived cells retaining spherical shape. Mutations in the peptidoglycan synthesis protein Pbp1A were identified as the main route for evolutionary rescue with genetic reconstructions demonstrating causality. Compensatory pbp1A mutations that targeted transpeptidase activity enhanced homogeneity of cell wall synthesis on lateral surfaces and restored cell size homeostasis. Mechanistic explanations require enhanced understanding of why deletion of mreB causes heterogeneity in cell wall synthesis. We conclude by presenting two testable hypotheses, one of which posits that heterogeneity stems from non-functional cell wall synthesis machinery, while the second posits that the machinery is functional, albeit stalled. Overall, our data provide support for the second hypothesis and draw attention to the importance of balance between transpeptidase and glycosyltransferase functions of peptidoglycan building enzymes for cell shape determination.

    1. Cell Biology
    Kaima Tsukada, Rikiya Imamura ... Mikio Shimada
    Research Article

    Polynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment to the gapped DNA structure found in single-strand DNA (ssDNA) nicks and/or gaps between Okazaki fragments (OFs) during DNA replication. T118A (alanine)-substituted PNKP-expressing cells exhibited an accumulation of ssDNA gaps in S phase and accelerated replication fork progression. Furthermore, PNKP is involved in poly (ADP-ribose) polymerase 1 (PARP1)-dependent replication gap filling as part of a backup pathway in the absence of OFs ligation. Altogether, our data suggest that CDK-mediated PNKP phosphorylation at T118 is important for its recruitment to ssDNA gaps to proceed with OFs ligation and its backup repairs via the gap-filling pathway to maintain genome stability.