1. Cell Biology
Download icon

CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels

  1. André B Goncalves
  2. Sarah K Hasselbalch
  3. Beinta B Joensen
  4. Sebastian Patzke
  5. Pernille Martens
  6. Signe K Ohlsen
  7. Mathieu Quinodoz
  8. Konstantinos Nikopoulos
  9. Reem Suleiman
  10. Magnus P Damso Jeppesen
  11. Catja Weiss
  12. Søren Tvorup Christensen
  13. Carlo Rivolta
  14. Jens S Andersen
  15. Pietro Farinelli  Is a corresponding author
  16. Lotte B Pedersen  Is a corresponding author
  1. University of Copenhagen, Denmark
  2. Oslo University Hospital, Norway
  3. University of Southern Denmark, Denmark
  4. University of Basel, Switzerland
  5. University of Lausanne, Switzerland
Research Article
  • Cited 0
  • Views 367
  • Annotations
Cite this article as: eLife 2021;10:e63731 doi: 10.7554/eLife.63731

Abstract

CEP78 is a centrosomal protein implicated in ciliogenesis and ciliary length control, and mutations in the CEP78 gene cause retinal cone-rod dystrophy associated with hearing loss. However, the mechanism by which CEP78 affects cilia formation is unknown. Based on a recently discovered disease-causing CEP78 p.L150S mutation, we identified the disease-relevant interactome of CEP78. We confirmed that CEP78 interacts with the EDD1-DYRK2-DDB1VPRBP E3 ubiquitin ligase complex, which is involved in CP110 ubiquitination and degradation, and identified a novel interaction between CEP78 and CEP350 that is weakened by the CEP78L150S mutation. We show that CEP350 promotes centrosomal recruitment and stability of CEP78, which in turn leads to centrosomal recruitment of EDD1. Consistently, cells lacking CEP78 display significantly increased cellular and centrosomal levels of CP110, and depletion of CP110 in CEP78-deficient cells restored ciliation frequency to normal. We propose that CEP78 functions downstream of CEP350 to promote ciliogenesis by negatively regulating CP110 levels via an EDD1-dependent mechanism.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figure 3A and Figure 7-figure supplement 2.

Article and author information

Author details

  1. André B Goncalves

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  2. Sarah K Hasselbalch

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  3. Beinta B Joensen

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  4. Sebastian Patzke

    Oslo University Hospital, Oslo, Norway
    Competing interests
    The authors declare that no competing interests exist.
  5. Pernille Martens

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  6. Signe K Ohlsen

    Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  7. Mathieu Quinodoz

    Institute of Molecular and Clinical Ophthalmology Basel (IOB); Department of Ophthalmology, University of Basel, Basel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  8. Konstantinos Nikopoulos

    Department of Computational Biology, University of Lausanne, Lausanne, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  9. Reem Suleiman

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  10. Magnus P Damso Jeppesen

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  11. Catja Weiss

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  12. Søren Tvorup Christensen

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5004-304X
  13. Carlo Rivolta

    Institute of Molecular and Clinical Ophthalmology Basel (IOB); Department of Ophthalmology, University of Basel, Basel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  14. Jens S Andersen

    Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
    Competing interests
    The authors declare that no competing interests exist.
  15. Pietro Farinelli

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    Pietro.Farinelli@twelve.bio
    Competing interests
    The authors declare that no competing interests exist.
  16. Lotte B Pedersen

    Department of Biology, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    lbpedersen@bio.ku.dk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9749-3758

Funding

Independent Research Fund Denmark (8020‐00162B)

  • Pietro Farinelli
  • Lotte B Pedersen

Carlsberg Foundation (CF18‐0294)

  • Lotte B Pedersen

Independent Research Fund Denmark (8021-00425A)

  • Jens S Andersen

Swiss National Science Foundation (176097)

  • Carlo Rivolta

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

Reviewing Editor

  1. Jens Lüders, Institute for Research in Biomedicine, Spain

Publication history

  1. Received: October 5, 2020
  2. Accepted: July 13, 2021
  3. Accepted Manuscript published: July 14, 2021 (version 1)

Copyright

© 2021, Goncalves 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

  • 367
    Page views
  • 84
    Downloads
  • 0
    Citations

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

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)

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

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

Further reading

    1. Cell Biology
    Alberto T Gatta et al.
    Research Article Updated

    Through membrane sealing and disassembly of spindle microtubules, the Endosomal Sorting Complex Required for Transport-III (ESCRT-III) machinery has emerged as a key player in the regeneration of a sealed nuclear envelope (NE) during mitotic exit, and in the repair of this organelle during interphase rupture. ESCRT-III assembly at the NE occurs transiently during mitotic (M) exit and is initiated when CHMP7, an ER-localised ESCRT-II/ESCRT-III hybrid protein, interacts with the Inner Nuclear Membrane (INM) protein LEM2. Whilst classical nucleocytoplasmic transport mechanisms have been proposed to separate LEM2 and CHMP7 during interphase, it is unclear how CHMP7 assembly is suppressed in mitosis when NE and ER identities are mixed. Here, we use live cell imaging and protein biochemistry to examine the biology of these proteins during M-exit. Firstly, we show that CHMP7 plays an important role in the dissolution of LEM2 clusters that form at the NE during M-exit. Secondly, we show that CDK1 phosphorylates CHMP7 upon M-entry at Ser3 and Ser441 and that this phosphorylation reduces CHMP7’s interaction with LEM2, limiting its assembly during M-phase. We show that spatiotemporal differences in the dephosphorylation of CHMP7 license its assembly at the NE during telophase, but restrict its assembly on the ER at this time. Without CDK1 phosphorylation, CHMP7 undergoes inappropriate assembly in the peripheral ER during M-exit, capturing LEM2 and downstream ESCRT-III components. Lastly, we establish that a microtubule network is dispensable for ESCRT-III assembly at the reforming nuclear envelope. These data identify a key cell-cycle control programme allowing ESCRT-III-dependent nuclear regeneration.

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Jingxiang Li et al.
    Research Article Updated

    Autophagy acts as a pivotal innate immune response against infection. Some virulence effectors subvert the host autophagic machinery to escape the surveillance of autophagy. The mechanism by which pathogens interact with host autophagy remains mostly unclear. However, traditional strategies often have difficulty identifying host proteins that interact with effectors due to the weak, dynamic, and transient nature of these interactions. Here, we found that Enteropathogenic Escherichia coli (EPEC) regulates autophagosome formation in host cells dependent on effector NleE. The 26S Proteasome Regulatory Subunit 10 (PSMD10) was identified as a direct interaction partner of NleE in living cells by employing genetically incorporated crosslinkers. Pairwise chemical crosslinking revealed that NleE interacts with the N-terminus of PSMD10. We demonstrated that PSMD10 homodimerization is necessary for its interaction with ATG7 and promotion of autophagy, but not necessary for PSMD10 interaction with ATG12. Therefore, NleE-mediated PSMD10 in monomeric state attenuates host autophagosome formation. Our study reveals the mechanism through which EPEC attenuates host autophagy activity.