Listeria monocytogenes requires cellular respiration for NAD+ regeneration and pathogenesis

  1. Rafael Rivera-Lugo
  2. David Deng
  3. Andrea Anaya-Sanchez
  4. Sara Tejedor-Sanz
  5. Eugene Tang
  6. Valeria M Reyes Ruiz
  7. Hans B Smith
  8. Denis V Titov
  9. John Demian Sauer
  10. Eric P Skaar
  11. Caroline M Ajo-Franklin
  12. Daniel A Portnoy
  13. Samuel H Light  Is a corresponding author
  1. University of California, Berkeley, United States
  2. Rice University, United States
  3. Vanderbilt University Medical Center, United States
  4. University of Wisconsin-Madison, United States
  5. University of Chicago, United States

Abstract

Cellular respiration is essential for multiple bacterial pathogens and a validated antibiotic target. In addition to driving oxidative phosphorylation, bacterial respiration has a variety of ancillary functions that obscure its contribution to pathogenesis. We find here that the intracellular pathogen Listeria monocytogenes encodes two respiratory pathways which are partially functionally redundant and indispensable for pathogenesis. Loss of respiration decreased NAD+ regeneration, but this could be specifically reversed by heterologous expression of a water-forming NADH oxidase (NOX). NOX expression fully rescued intracellular growth defects and increased L. monocytogenes loads >1,000-fold in a mouse infection model. Consistent with NAD+ regeneration maintaining L. monocytogenes viability and enabling immune evasion, a respiration-deficient strain exhibited elevated bacteriolysis within the host cytosol and NOX expression rescued this phenotype. These studies show that NAD+ regeneration represents a major role of L. monocytogenes respiration and highlight the nuanced relationship between bacterial metabolism, physiology, and pathogenesis.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Rafael Rivera-Lugo

    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2346-2297
  2. David Deng

    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
  3. Andrea Anaya-Sanchez

    Graduate Group in Microbiology, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
  4. Sara Tejedor-Sanz

    Department of Biosciences, Rice University, Houston, United States
    Competing interests
    No competing interests declared.
  5. Eugene Tang

    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
  6. Valeria M Reyes Ruiz

    Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, United States
    Competing interests
    No competing interests declared.
  7. Hans B Smith

    Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, United States
    Competing interests
    No competing interests declared.
  8. Denis V Titov

    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
    Competing interests
    Denis V Titov, is a co-inventor on a filed patent describing the use of NOX. (US Patent App. 15/749,218).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5677-0651
  9. John Demian Sauer

    Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9367-794X
  10. Eric P Skaar

    Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, United States
    Competing interests
    No competing interests declared.
  11. Caroline M Ajo-Franklin

    Department of Biosciences, Rice University, Houston, United States
    Competing interests
    No competing interests declared.
  12. Daniel A Portnoy

    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
  13. Samuel H Light

    Department of Microbiology, University of Chicago, Chicago, United States
    For correspondence
    samlight@uchicago.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8074-1348

Funding

National Institutes of Health (T32GM007215)

  • Hans B Smith

Searle Scholars Program

  • Samuel H Light

National Institutes of Health (R01AI137070)

  • John Demian Sauer

National Institutes of Health (R01AI073843)

  • Eric P Skaar

National Institutes of Health (R01AI073843)

  • Eric P Skaar

National Institutes of Health (1P01AI063302)

  • Daniel A Portnoy

National Institutes of Health (1R01AI27655)

  • Daniel A Portnoy

National Institutes of Health (K22AI144031)

  • Samuel H Light

National Academies of Sciences, Engineering, and Medicine (Ford Foundation Fellowship)

  • Rafael Rivera-Lugo

University of California (Dissertation-Year Fellowship)

  • Rafael Rivera-Lugo

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

Ethics

Animal experimentation: All animal work was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Protocols were reviewed and approved by the Animal Care and Use Committee at the University of California, Berkeley (AUP 2016-05-8811).

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 3,061
    views
  • 449
    downloads
  • 19
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Rafael Rivera-Lugo
  2. David Deng
  3. Andrea Anaya-Sanchez
  4. Sara Tejedor-Sanz
  5. Eugene Tang
  6. Valeria M Reyes Ruiz
  7. Hans B Smith
  8. Denis V Titov
  9. John Demian Sauer
  10. Eric P Skaar
  11. Caroline M Ajo-Franklin
  12. Daniel A Portnoy
  13. Samuel H Light
(2022)
Listeria monocytogenes requires cellular respiration for NAD+ regeneration and pathogenesis
eLife 11:e75424.
https://doi.org/10.7554/eLife.75424

Share this article

https://doi.org/10.7554/eLife.75424

Further reading

    1. Biochemistry and Chemical Biology
    Nelson García-Vázquez, Tania J González-Robles ... Michele Pagano
    Research Article

    In healthy cells, cyclin D1 is expressed during the G1 phase of the cell cycle, where it activates CDK4 and CDK6. Its dysregulation is a well-established oncogenic driver in numerous human cancers. The cancer-related function of cyclin D1 has been primarily studied by focusing on the phosphorylation of the retinoblastoma (RB) gene product. Here, using an integrative approach combining bioinformatic analyses and biochemical experiments, we show that GTSE1 (G-Two and S phases expressed protein 1), a protein positively regulating cell cycle progression, is a previously unrecognized substrate of cyclin D1–CDK4/6 in tumor cells overexpressing cyclin D1 during G1 and subsequent phases. The phosphorylation of GTSE1 mediated by cyclin D1–CDK4/6 inhibits GTSE1 degradation, leading to high levels of GTSE1 across all cell cycle phases. Functionally, the phosphorylation of GTSE1 promotes cellular proliferation and is associated with poor prognosis within a pan-cancer cohort. Our findings provide insights into cyclin D1’s role in cell cycle control and oncogenesis beyond RB phosphorylation.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Mai Nguyen, Elda Bauda ... Cecile Morlot
    Research Article

    Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.