1. Biochemistry and Chemical Biology
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Fine interaction profiling of VemP and mechanisms responsible for its translocation-coupled arrest-cancelation

  1. Ryoji Miyazaki
  2. Yoshinori Akiyama
  3. Hiroyuki Mori  Is a corresponding author
  1. Kyoto University, Japan
Research Article
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Cite this article as: eLife 2020;9:e62623 doi: 10.7554/eLife.62623

Abstract

Bacterial cells utilize monitoring substrates, which undergo force-sensitive translation elongation arrest, to feedback-regulate a Sec-related gene. Vibrio alginolyticus VemP controls the expression of SecD/F that stimulates a late step of translocation by undergoing export-regulated elongation arrest. Here, we attempted at delineating the pathway of the VemP nascent-chain interaction with Sec-related factors, and identified the signal recognition particle (SRP) and PpiD (a membrane-anchored periplasmic chaperone) in addition to other translocon components and a ribosomal protein as interacting partners. Our results showed that SRP is required for the membrane-targeting of VemP, whereas PpiD acts cooperatively with SecD/F in the translocation and arrest-cancelation of VemP. We also identified the conserved Arg-85 residue of VemP as a crucial element that confers PpiD-dependence to VemP and plays an essential role in the regulated arrest-cancelation. We propose a scheme of the arrest-cancelation processes of VemP, which likely monitors late steps in the protein translocation pathway.

Data availability

All data generated and analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 2, 3 and 4, Figure 2-figure supplements 1, 3 and 4 and Figure 5-figure supplements 1 and 2.

Article and author information

Author details

  1. Ryoji Miyazaki

    Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  2. Yoshinori Akiyama

    Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4483-5408
  3. Hiroyuki Mori

    Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan
    For correspondence
    hiromori@infront.kyoto-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0429-1269

Funding

Japan Society for the Promotion of Science (18H06047)

  • Ryoji Miyazaki

Japan Society for the Promotion of Science (19K21179)

  • Ryoji Miyazaki

Japan Society for the Promotion of Science (20K15715)

  • Ryoji Miyazaki

Japan Society for the Promotion of Science (15H01532)

  • Yoshinori Akiyama

Japan Society for the Promotion of Science (18H02404)

  • Yoshinori Akiyama

Japan Society for the Promotion of Science (17H05666)

  • Hiroyuki Mori

Japan Society for the Promotion of Science (17H05879)

  • Hiroyuki Mori

Japan Society for the Promotion of Science (17K07334)

  • Hiroyuki Mori

Japan Society for the Promotion of Science (20K06556)

  • Hiroyuki Mori

The funders had no role in study design, data collection and interpretation.

Reviewing Editor

  1. Suzanne R Pfeffer, Stanford University School of Medicine, United States

Publication history

  1. Received: August 31, 2020
  2. Accepted: December 11, 2020
  3. Accepted Manuscript published: December 15, 2020 (version 1)
  4. Version of Record published: January 8, 2021 (version 2)
  5. Version of Record updated: January 13, 2021 (version 3)

Copyright

© 2020, Miyazaki 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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