Bardet-Biedl Syndrome 3 protein promotes ciliary exit of the signaling protein phospholipase D via the BBSome

  1. Yan-Xia Liu
  2. Bin Xue
  3. Wei-Yue Sun
  4. Jenna L Wingfield
  5. Jun Sun
  6. Mingfu Wu
  7. Karl F Lechtreck
  8. Zhenlong Wu
  9. Zhen-Chuan Fan  Is a corresponding author
  1. Tianjin University of Science and Technology, China
  2. University of Georgia, United States
  3. Albany Medical College, United States
  4. China Agricultural University, China

Abstract

Certain ciliary signaling proteins couple with the BBSome, a conserved complex of Bardet-Biedl syndrome (BBS) proteins, to load onto retrograde intraflagellar transport (IFT) trains for their removal out of cilia in Chlamydomonas reinhardtii. Here, we show that loss of the Arf-like 6 (ARL6) GTPase BBS3 causes the signaling protein phospholipase D (PLD) to accumulate in cilia. Upon targeting to the basal body, BBSomes enter and cycle through cilia via IFT, while BBS3 in a GTP-bound state separates from BBSomes, associates with the membrane, and translocates from the basal body to cilia by diffusion. Upon arriving at the ciliary tip, GTP-bound BBS3 binds and recruits BBSomes to the ciliary membrane for interacting with PLD, thus making the PLD-laden BBSomes available to load onto retrograde IFT trains for ciliary exit. Therefore, BBS3 promotes PLD exit from cilia via the BBSome providing a regulatory mechanism for ciliary signaling protein removal out of cilia.

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. Yan-Xia Liu

    Institute of Health Biotechnology, Tianjin University of Science and Technology, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Bin Xue

    Institute of Health Biotechnology, Tianjin University of Science and Technology, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Wei-Yue Sun

    Institute of Health Biotechnology, Tianjin University of Science and Technology, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Jenna L Wingfield

    Department of Cellular Biology, University of Georgia, Athens, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Jun Sun

    Institute of Health Biotechnology, Tianjin University of Science and Technology, Tianjin, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Mingfu Wu

    Department of Molecular and Cellular Physiology, Albany Medical College, Albany, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Karl F Lechtreck

    Department of Cellular Biology, University of Georgia, Athens, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Zhenlong Wu

    State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Zhen-Chuan Fan

    Institute of Health Biotechnology, Tianjin University of Science and Technology, Tianjin, China
    For correspondence
    fanzhen@tust.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3007-4281

Funding

International Center for Genetic Engineering and Biotechnology (CRP/CHN15-01)

  • Zhen-Chuan Fan

National Natural Science Foundation of China (41876134)

  • Jun Sun

National Institutes of Health (GM110413)

  • Karl F Lechtreck

National Natural Science Foundation of China (32070698)

  • Zhen-Chuan Fan

Tianjin Municipal Science and Technology Bureau (19PTSYJC00050)

  • Zhen-Chuan Fan

Tianjin Municipal Science and Technology Bureau (18JCZDJC34100)

  • Zhen-Chuan Fan

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

Reviewing Editor

  1. Junmin Pan, Tsinghua University, China

Version history

  1. Received: May 20, 2020
  2. Accepted: February 13, 2021
  3. Accepted Manuscript published: February 15, 2021 (version 1)
  4. Version of Record published: March 16, 2021 (version 2)

Copyright

© 2021, Liu 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

  • 1,362
    Page views
  • 196
    Downloads
  • 19
    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)

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. Yan-Xia Liu
  2. Bin Xue
  3. Wei-Yue Sun
  4. Jenna L Wingfield
  5. Jun Sun
  6. Mingfu Wu
  7. Karl F Lechtreck
  8. Zhenlong Wu
  9. Zhen-Chuan Fan
(2021)
Bardet-Biedl Syndrome 3 protein promotes ciliary exit of the signaling protein phospholipase D via the BBSome
eLife 10:e59119.
https://doi.org/10.7554/eLife.59119

Further reading

    1. Cell Biology
    2. Microbiology and Infectious Disease
    Heledd Davies, Hugo Belda ... Moritz Treeck
    Tools and Resources

    Reverse genetics is key to understanding protein function, but the mechanistic connection between a gene of interest and the observed phenotype is not always clear. Here we describe the use of proximity labeling using TurboID and site-specific quantification of biotinylated peptides to measure changes to the local protein environment of selected targets upon perturbation. We apply this technique, which we call PerTurboID, to understand how the P. falciparum exported kinase, FIKK4.1, regulates the function of the major virulence factor of the malaria causing parasite, PfEMP1. We generated independent TurboID fusions of 2 proteins that are predicted substrates of FIKK4.1 in a FIKK4.1 conditional KO parasite line. Comparing the abundance of site-specific biotinylated peptides between wildtype and kinase deletion lines reveals the differential accessibility of proteins to biotinylation, indicating changes to localization, protein-protein interactions, or protein structure which are mediated by FIKK4.1 activity. We further show that FIKK4.1 is likely the only FIKK kinase that controls surface levels of PfEMP1, but not other surface antigens, on the infected red blood cell under standard culture conditions. We believe PerTurboID is broadly applicable to study the impact of genetic or environmental perturbation on a selected cellular niche.

    1. Cell Biology
    Bo Wang, Zheyong Liang ... Peijun Liu
    Research Article

    The primary cilium plays important roles in regulating cell differentiation, signal transduction, and tissue organization. Dysfunction of the primary cilium can lead to ciliopathies and cancer. The formation and organization of the primary cilium are highly associated with cell polarity proteins, such as the apical polarity protein CRB3. However, the molecular mechanisms by which CRB3 regulates ciliogenesis and the location of CRB3 remain unknown. Here, we show that CRB3, as a navigator, regulates vesicle trafficking in γ-tubulin ring complex (γTuRC) assembly during ciliogenesis and cilium-related Hh and Wnt signaling pathways in tumorigenesis. Crb3 knockout mice display severe defects of the primary cilium in the mammary ductal lumen and renal tubule, while mammary epithelial-specific Crb3 knockout mice exhibit the promotion of ductal epithelial hyperplasia and tumorigenesis. CRB3 is essential for lumen formation and ciliary assembly in the mammary epithelium. We demonstrate that CRB3 localizes to the basal body and that CRB3 trafficking is mediated by Rab11-positive endosomes. Significantly, CRB3 interacts with Rab11 to navigate GCP6/Rab11 trafficking vesicles to CEP290, resulting in intact γTuRC assembly. In addition, CRB3-depleted cells are unresponsive to the activation of the Hh signaling pathway, while CRB3 regulates the Wnt signaling pathway. Therefore, our studies reveal the molecular mechanisms by which CRB3 recognizes Rab11-positive endosomes to facilitate ciliogenesis and regulates cilium-related signaling pathways in tumorigenesis.