1. Biochemistry and Chemical Biology
  2. Microbiology and Infectious Disease
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Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria

  1. Alam García-Heredia
  2. Amol A Pohane
  3. Emily S Melzer
  4. Caleb R Carr
  5. Taylor J Fiolek  Is a corresponding author
  6. Sarah R Rundell
  7. Hoong Chuin Lim
  8. Jeffrey C Wagner
  9. Yasu S Morita
  10. Benjamin M Swarts
  11. M Sloan Siegrist  Is a corresponding author
  1. University of Massachusetts, United States
  2. Central Michigan University, United States
  3. Harvard Medical School, United States
  4. Harvard TH Chan School of Public Health, United States
Research Article
  • Cited 7
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Cite this article as: eLife 2018;7:e37243 doi: 10.7554/eLife.37243

Abstract

Rod-shaped mycobacteria expand from their poles, yet d-amino acid probes label cell wall peptidoglycan in this genus at both the poles and sidewall. We sought to clarify the metabolic fates of these probes. Monopeptide incorporation was decreased by antibiotics that block peptidoglycan synthesis or l,d-transpeptidation and in an l,d-transpeptidase mutant. Dipeptides complemented defects in d-alanine synthesis or ligation and were present in lipid-linked peptidoglycan precursors. Characterizing probe uptake pathways allowed us to localize peptidoglycan metabolism with precision: monopeptide-marked l,d-transpeptidase remodeling and dipeptide-marked synthesis were coincident with mycomembrane metabolism at the poles, septum and sidewall. Fluorescent pencillin-marked d,d-transpeptidation around the cell perimeter further suggested that the mycobacterial sidewall is a site of cell wall assembly. While polar peptidoglycan synthesis was associated with cell elongation, sidewall synthesis responded to cell wall damage. Peptidoglycan editing along the sidewall may support cell wall robustness in pole-growing mycobacteria.

Article and author information

Author details

  1. Alam García-Heredia

    Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9573-4087
  2. Amol A Pohane

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

    Department of Microbiology, University of Massachusetts, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Caleb R Carr

    Department of Microbiology, University of Massachusetts, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Taylor J Fiolek

    Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, United States
    For correspondence
    fiole1tj@cmich.edu
    Competing interests
    The authors declare that no competing interests exist.
  6. Sarah R Rundell

    Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Hoong Chuin Lim

    Department of Microbiology and Immunobiology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Jeffrey C Wagner

    Department of Immunology and Infectious Diseases, Harvard TH Chan School of Public Health, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Yasu S Morita

    Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4514-9242
  10. Benjamin M Swarts

    Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. M Sloan Siegrist

    Molecular and Cellular Biology Graduate Program, University of Massachusetts, 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 Institutes of Health (New Innovator Award DP2 AI138238)

  • M Sloan Siegrist

National Science Foundation (CAREER 1654408)

  • Benjamin M Swarts

Simons Foundation (Life Sciences Research Foundation Fellowship)

  • Hoong Chuin Lim

Research Corporation for Science Advancement (Cottrell College Science Award 22525)

  • Benjamin M Swarts

National Institutes of Health (U01CA221230)

  • M Sloan Siegrist

National Institutes of Health (Training Grant T32 GM008515)

  • Emily S Melzer

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

Reviewing Editor

  1. Bavesh D Kana, University of the Witwatersrand, South Africa

Publication history

  1. Received: April 8, 2018
  2. Accepted: September 9, 2018
  3. Accepted Manuscript published: September 10, 2018 (version 1)
  4. Version of Record published: October 16, 2018 (version 2)
  5. Version of Record updated: May 28, 2019 (version 3)

Copyright

© 2018, García-Heredia 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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