Genetically incorporated crosslinkers reveal NleE attenuates host autophagy dependent on PSMD10

Abstract

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.

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

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Author details

  1. Jingxiang Li

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Shupan Guo

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Li Zhou

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Fangni Chai

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Qi Sun

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Pan Li

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Li Gao

    West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Lunzhi Dai

    West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Xiaoxiao Ouyang

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Zhihui Zhou

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  11. Wei Cheng

    West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  12. Shiqian Qi

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  13. Kefeng Lu

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    Competing interests
    The authors declare that no competing interests exist.
  14. Haiyan Ren

    Division of Respiratory and Critical Care Medicine, State Key Laboratory of Biotherapy, West China Hospital of Sichuan University and Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu, China
    For correspondence
    hyren@scu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9995-3255

Funding

National Key Research and Development Program of China (2018YFC1002802)

  • Haiyan Ren

National Key Research and Development Program of China (2018YFA0109200)

  • Haiyan Ren

National Natural Science Foundation of China (31872727)

  • Haiyan Ren

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

Reviewing Editor

  1. Ivan Dikic, Institute of Biochemistry II, Germany

Publication history

  1. Received: April 1, 2021
  2. Accepted: July 13, 2021
  3. Accepted Manuscript published: July 13, 2021 (version 1)
  4. Version of Record published: July 30, 2021 (version 2)

Copyright

© 2021, Li 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.

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  1. Jingxiang Li
  2. Shupan Guo
  3. Li Zhou
  4. Fangni Chai
  5. Qi Sun
  6. Pan Li
  7. Li Gao
  8. Lunzhi Dai
  9. Xiaoxiao Ouyang
  10. Zhihui Zhou
  11. Wei Cheng
  12. Shiqian Qi
  13. Kefeng Lu
  14. Haiyan Ren
(2021)
Genetically incorporated crosslinkers reveal NleE attenuates host autophagy dependent on PSMD10
eLife 10:e69047.
https://doi.org/10.7554/eLife.69047
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