A cell wall synthase accelerates plasma membrane partitioning in mycobacteria

  1. Takehiro Kado
  2. Zarina Akbary
  3. Daisuke Motooka
  4. Ian L Sparks
  5. Emily S Melzer
  6. Shota Nakamura
  7. Enrique R Rojas
  8. Yasu S Morita  Is a corresponding author
  9. M Sloan Siegrist  Is a corresponding author
  1. University of Massachusetts Amherst, United States
  2. New York University, United States
  3. Osaka University, Japan

Abstract

Lateral partitioning of proteins and lipids shapes membrane function. In model membranes, partitioning can be influenced both by bilayer-intrinsic factors like molecular composition and by bilayer-extrinsic factors such as interactions with other membranes and solid supports. While cellular membranes can departition in response to bilayer-intrinsic or -extrinsic disruptions, the mechanisms by which they partition de novo are largely unknown. The plasma membrane of Mycobacterium smegmatis spatially and biochemically departitions in response to the fluidizing agent benzyl alcohol, then repartitions upon fluidizer washout. By screening for mutants that are sensitive to benzyl alcohol, we show that the bifunctional cell wall synthase PonA2 promotes membrane partitioning and cell growth during recovery from benzyl alcohol exposure. PonA2's role in membrane repartitioning and regrowth depends solely on its conserved transglycosylase domain. Active cell wall polymerization promotes de novo membrane partitioning and the completed cell wall polymer helps to maintain membrane partitioning. Our work highlights the complexity of membrane-cell wall interactions and establishes a facile model system for departitioning and repartitioning cellular membranes.

Data availability

Sequencing data have been deposited in NIH SRA under accession codes PRJNA976743.All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for all Figures and figure supplements.

The following data sets were generated

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Author details

  1. Takehiro Kado

    Department of Microbiology, University of Massachusetts Amherst, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5419-6123
  2. Zarina Akbary

    Department of Biology, New York University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Daisuke Motooka

    Department of Infection Metagenomics, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.
  4. Ian L Sparks

    Department of Microbiology, University of Massachusetts Amherst, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Emily S Melzer

    Department of Microbiology, University of Massachusetts Amherst, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Shota Nakamura

    Department of Infection Metagenomics, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.
  7. Enrique R Rojas

    Department of Biology, New York University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3388-2794
  8. Yasu S Morita

    Department of Microbiology, University of Massachusetts Amherst, Amherst, United States
    For correspondence
    ymorita@umass.edu
    Competing interests
    The authors declare that no competing interests exist.
  9. M Sloan Siegrist

    Molecular and Cellular Graduate Program, University of Massachusetts Amherst, Amherst, United States
    For correspondence
    siegrist@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8232-3246

Funding

National Institute of Allergy and Infectious Diseases (R21 AI144748)

  • Yasu S Morita

National Institute of Allergy and Infectious Diseases (R21 AI144748)

  • M Sloan Siegrist

National Institute of Allergy and Infectious Diseases (DP2 AI138238)

  • M Sloan Siegrist

National Institute of Allergy and Infectious Diseases (R03 AI140259-01)

  • Yasu S Morita

National Institute of General Medical Sciences (R35GM143057)

  • Enrique R Rojas

National Institute of General Medical Sciences (T32 GM008515)

  • Emily S Melzer

Uehara Memorial Foundation

  • Takehiro Kado

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

Reviewing Editor

  1. Petra Anne Levin, Washington University in St. Louis, United States

Version history

  1. Preprint posted: June 14, 2022 (view preprint)
  2. Received: July 16, 2022
  3. Accepted: September 2, 2023
  4. Accepted Manuscript published: September 4, 2023 (version 1)
  5. Version of Record published: October 3, 2023 (version 2)

Copyright

© 2023, Kado 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. Takehiro Kado
  2. Zarina Akbary
  3. Daisuke Motooka
  4. Ian L Sparks
  5. Emily S Melzer
  6. Shota Nakamura
  7. Enrique R Rojas
  8. Yasu S Morita
  9. M Sloan Siegrist
(2023)
A cell wall synthase accelerates plasma membrane partitioning in mycobacteria
eLife 12:e81924.
https://doi.org/10.7554/eLife.81924

Share this article

https://doi.org/10.7554/eLife.81924

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