The transpeptidase PBP2 governs initial localization and activity of the major cell-wall synthesis machinery in E. coli

Abstract

Bacterial shape is physically determined by the peptidoglycan cell wall. The cell-wall-synthesis machinery responsible for rod shape in Escherichia coli is the processive 'Rod complex'. Previously, cytoplasmic MreB filaments were thought to govern formation and localization of Rod complexes based on local cell-envelope curvature. Using single-particle tracking of the transpeptidase and Rod-complex component PBP2, we found that PBP2 binds to a substrate different from MreB. Depletion and localization experiments of other putative Rod-complex components provide evidence that none of those provide the sole rate-limiting substrate for PBP2 binding. Consistently, we found only weak correlations between MreB and envelope curvature in the cylindrical part of cells. Residual correlations do not require curvature-based Rod-complex initiation but can be attributed to persistent rotational motion. We therefore speculate that the local cell-wall architecture provides the cue for Rod-complex initiation, either through direct binding by PBP2 or through an unknown intermediate.

Data availability

All data generated or analysed during this study are included in supplemental datasets provided for each figure. Source data, specifically raw tracks, are provided as Source Data File (one file with x-, y- coordinates and track identifier per replicate).

Article and author information

Author details

  1. Gizem Özbaykal

    Microbial Morphogenesis and Growth Laboratory, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Eva Wollrab

    Department of Microbiology, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Francois Simon

    Department of Microbiology, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Antoine Vigouroux

    Microbial Morphogenesis and Growth Laboratory, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8398-5073
  5. Baptiste Cordier

    Microbial Morphogenesis and Growth Laboratory, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Andrey Aristov

    Microbial Morphogenesis and Growth Laboratory, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Thibault Chaze

    Proteomics Platform, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  8. Mariette Matondo

    Proteomics Platform, Institut Pasteur, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  9. Sven van Teeffelen

    Microbial Morphogenesis and Growth Laboratory, Institut Pasteur, Paris, France
    For correspondence
    sven.vanteeffelen@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0877-1294

Funding

H2020 European Research Council (679980)

  • Sven van Teeffelen

Agence Nationale de la Recherche (ANR-10-LABX-62-IBEID)

  • Sven van Teeffelen

Volkswagen Foundation

  • Sven van Teeffelen

Mairie de Paris

  • Sven van Teeffelen

Prestige Postdoctoral Fellowship

  • Eva Wollrab

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

Reviewing Editor

  1. Tâm Mignot, CNRS-Aix Marseille University, France

Publication history

  1. Received: July 28, 2019
  2. Accepted: February 19, 2020
  3. Accepted Manuscript published: February 20, 2020 (version 1)
  4. Version of Record published: March 23, 2020 (version 2)

Copyright

© 2020, Özbaykal 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. Gizem Özbaykal
  2. Eva Wollrab
  3. Francois Simon
  4. Antoine Vigouroux
  5. Baptiste Cordier
  6. Andrey Aristov
  7. Thibault Chaze
  8. Mariette Matondo
  9. Sven van Teeffelen
(2020)
The transpeptidase PBP2 governs initial localization and activity of the major cell-wall synthesis machinery in E. coli
eLife 9:e50629.
https://doi.org/10.7554/eLife.50629

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