Group II truncated haemoglobin YjbI prevents reactive oxygen species-induced protein aggregation in Bacillus subtilis

  1. Takeshi Imai  Is a corresponding author
  2. Ryuta Tobe
  3. Koji Honda
  4. Mai Tanaka
  5. Jun Kawamoto
  6. Hisaaki Mihara  Is a corresponding author
  1. Hyogo Prefectural Institute of Technology, Japan
  2. Ritsumeikan University, Japan
  3. Kyoto University, Japan

Abstract

Oxidative stress–mediated formation of protein hydroperoxides can induce irreversible fragmentation of the peptide backbone and accumulation of cross-linked protein aggregates, leading to cellular toxicity, dysfunction, and death. However, how bacteria protect themselves from damages caused by protein hydroperoxidation is unknown. Here we show that YjbI, a group II truncated haemoglobin from Bacillus subtilis, prevents oxidative aggregation of cell-surface proteins by its protein hydroperoxide peroxidase-like activity, which removes hydroperoxide groups from oxidised proteins. Disruption of the yjbI gene in B. subtilis lowered biofilm water repellence, which associated with the cross-linked aggregation of the biofilm matrix protein TasA. YjbI was localised to the cell surface or the biofilm matrix, and the sensitivity of planktonically grown cells to generators of reactive oxygen species was significantly increased upon yjbI disruption, suggesting that YjbI pleiotropically protects labile cell-surface proteins from oxidative damage. YjbI removed hydroperoxide residues from the model oxidized protein substrate bovine serum albumin and biofilm component TasA, preventing oxidative aggregation in vitro. Furthermore, the replacement of Tyr63 near the haem of YjbI with phenylalanine resulted in the loss of its protein peroxidase-like activity, and the mutant gene failed to rescue biofilm water repellency and resistance to oxidative stress induced by hypochlorous acid in the yjbI-deficient strain. These findings provide new insights into the role of truncated haemoglobin and the importance of hydroperoxide removal from proteins in the survival of aerobic bacteria.

Data availability

All data is available within the text, figures, and tables of the manuscript. Source data files have been provided for Figures 2, 3, Figure 2-figure supplement 1, Figure 3-figure supplement 1 and Figure 4-figure supplement 1.

Article and author information

Author details

  1. Takeshi Imai

    Hyogo Prefectural Institute of Technology, Hyogo, Japan
    For correspondence
    imai@hyogo-kg.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7387-6212
  2. Ryuta Tobe

    Department of Biotechnology, Ritsumeikan University, Shiga, Japan
    Competing interests
    The authors declare that no competing interests exist.
  3. Koji Honda

    Hyogo Prefectural Institute of Technology, Hyogo, Japan
    Competing interests
    The authors declare that no competing interests exist.
  4. Mai Tanaka

    Department of Biotechnology, Ritsumeikan University, Shiga, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Jun Kawamoto

    Institute for Chemical Research, Kyoto University, Kyoto, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Hisaaki Mihara

    Department of Biotechnology, Ritsumeikan University, Shiga, Japan
    For correspondence
    mihara@fc.ritsumei.ac.jp
    Competing interests
    The authors declare that no competing interests exist.

Funding

Japan Society for the Promotion of Science (18K14383)

  • Takeshi Imai

Japan Society for the Promotion of Science (20K15446)

  • Takeshi Imai

Ritsumeikan Global Innovation Research Organization, Ritsumeikan University

  • Hisaaki Mihara

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

Reviewing Editor

  1. Agnese Seminara, University of Genoa, Italy

Publication history

  1. Received: May 18, 2021
  2. Preprint posted: May 28, 2021 (view preprint)
  3. Accepted: September 19, 2022
  4. Accepted Manuscript published: September 20, 2022 (version 1)

Copyright

© 2022, Imai 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. Takeshi Imai
  2. Ryuta Tobe
  3. Koji Honda
  4. Mai Tanaka
  5. Jun Kawamoto
  6. Hisaaki Mihara
(2022)
Group II truncated haemoglobin YjbI prevents reactive oxygen species-induced protein aggregation in Bacillus subtilis
eLife 11:e70467.
https://doi.org/10.7554/eLife.70467
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