1. Microbiology and Infectious Disease
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Redox controls RecA protein activity via reversible oxidation of its methionine residues

  1. Camille Henry
  2. Laurent Loiseau
  3. Alexandra Vergnes
  4. Didier Vertommen
  5. Angela Mérida-Floriano
  6. Sindhu Chitteni-Pattu
  7. Elizabeth A Wood
  8. Josep Casadesús
  9. Michael M Cox
  10. Frédéric Barras  Is a corresponding author
  11. Benjamin Ezraty  Is a corresponding author
  1. University of Wisconsin-Madison, United States
  2. Aix-Marseille University, CNRS, France
  3. Université Catholique de Louvain, Belgium
  4. Universidad de Sevilla, Spain
  5. Institut Pasteur, France
Research Article
  • Cited 2
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Cite this article as: eLife 2021;10:e63747 doi: 10.7554/eLife.63747

Abstract

Reactive oxygen species (ROS) cause damage to DNA and proteins. Here we report that the RecA recombinase is itself oxidized by ROS. Genetic and biochemical analyses revealed that oxidation of RecA altered its DNA repair and DNA recombination activities. Mass spectrometry analysis showed that exposure to ROS converted 4 out of 9 Met residues of RecA to methionine sulfoxide. Mimicking oxidation of Met35 by changing it for Gln caused complete loss of function whereas mimicking oxidation of Met164 resulted in constitutive SOS activation and loss of recombination activity. Yet, all ROS-induced alterations of RecA activity were suppressed by methionine sulfoxide reductases MsrA and MsrB. These findings indicate that under oxidative stress, MsrA/B is needed for RecA homeostasis control. The implication is that, besides damaging DNA structure directly, ROS prevent repair of DNA damage by hampering RecA activity.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided in Dryad (doi:10.5061/dryad.zpc866t78).

The following data sets were generated

Article and author information

Author details

  1. Camille Henry

    Departement of Biochemistry, University of Wisconsin-Madison, Madison, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Laurent Loiseau

    Laboratoire de Chimie Bactérienne, Aix-Marseille University, CNRS, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Alexandra Vergnes

    Laboratoire de Chimie Bactérienne, Aix-Marseille University, CNRS, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Didier Vertommen

    Protein Phosphorylation Unit, de Duve Institute,, Université Catholique de Louvain, Brussels, Belgium
    Competing interests
    The authors declare that no competing interests exist.
  5. Angela Mérida-Floriano

    Departamento de Genética, Universidad de Sevilla, Sevilla, Spain
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9650-7759
  6. Sindhu Chitteni-Pattu

    Departement of Biochemistry, University of Wisconsin-Madison, Madison, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Elizabeth A Wood

    Department of Biochemistry, University of Wisconsin-Madison, Madison, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Josep Casadesús

    Departamento de Genética, Universidad de Sevilla, Sevilla, Spain
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2308-293X
  9. Michael M Cox

    Department of Biochemistry, University of Wisconsin-Madison, Madison, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3606-5722
  10. Frédéric Barras

    Institut Pasteur, Paris, France
    For correspondence
    frederic.barras@pasteur.fr
    Competing interests
    The authors declare that no competing interests exist.
  11. Benjamin Ezraty

    Laboratoire de Chimie Bactérienne, Aix-Marseille University, CNRS, Marseille, France
    For correspondence
    ezraty@imm.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3818-6907

Funding

Agence Nationale de la Recherche (ANR-METOXIC)

  • Benjamin Ezraty

Centre National de la Recherche Scientifique (PICS-PROTOX)

  • Benjamin Ezraty

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

  • Frédéric Barras

Fondation pour la Recherche Médicale

  • Camille Henry

Aix-Marseille Université (AMidex)

  • Camille Henry

National Institute of General Medical Sciences (GM32335)

  • Michael M Cox

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

Reviewing Editor

  1. Stephen C Kowalczykowski, University of California, Davis, United States

Publication history

  1. Received: October 6, 2020
  2. Accepted: February 18, 2021
  3. Accepted Manuscript published: February 19, 2021 (version 1)
  4. Accepted Manuscript updated: February 23, 2021 (version 2)
  5. Version of Record published: March 8, 2021 (version 3)

Copyright

© 2021, Henry 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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Further reading

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    Alexander O Pasternak et al.
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    Background:

    It remains unclear whether combination antiretroviral therapy (ART) regimens differ in their ability to fully suppress human immunodeficiency virus (HIV) replication. Here, we report the results of two cross-sectional studies that compared levels of cell-associated (CA) HIV markers between individuals receiving suppressive ART containing either a non-nucleoside reverse transcriptase inhibitor (NNRTI) or a protease inhibitor (PI).

    Methods:

    CA HIV unspliced RNA and total HIV DNA were quantified in two cohorts (n = 100, n = 124) of individuals treated with triple ART regimens consisting of two nucleoside reverse transcriptase inhibitors (NRTIs) plus either an NNRTI or a PI. To compare CA HIV RNA and DNA levels between the regimens, we built multivariable models adjusting for age, gender, current and nadir CD4+ count, plasma viral load zenith, duration of virological suppression, NRTI backbone composition, low-level plasma HIV RNA detectability, and electronically measured adherence to ART.

    Results:

    In both cohorts, levels of CA HIV RNA and DNA strongly correlated (rho = 0.70 and rho = 0.54) and both markers were lower in NNRTI-treated than in PI-treated individuals. In the multivariable analysis, CA RNA in both cohorts remained significantly reduced in NNRTI-treated individuals (padj = 0.02 in both cohorts), with a similar but weaker association between the ART regimen and total HIV DNA (padj = 0.048 and padj = 0.10). No differences in CA HIV RNA or DNA levels were observed between individual NNRTIs or individual PIs, but CA HIV RNA was lower in individuals treated with either nevirapine or efavirenz, compared to PI-treated individuals.

    Conclusions:

    All current classes of antiretroviral drugs only prevent infection of new cells but do not inhibit HIV RNA transcription in long-lived reservoir cells. Therefore, these differences in CA HIV RNA and DNA levels by treatment regimen suggest that NNRTIs are more potent in suppressing HIV residual replication than PIs, which may result in a smaller viral reservoir size.

    Funding:

    This work was supported by ZonMw (09120011910035) and FP7 Health (305522).

    1. Evolutionary Biology
    2. Microbiology and Infectious Disease
    Emily R Ebel et al.
    Research Article

    The replication of Plasmodium falciparum parasites within red blood cells (RBCs) causes severe disease in humans, especially in Africa. Deleterious alleles like hemoglobin S are well-known to confer strong resistance to malaria, but the effects of common RBC variation are largely undetermined. Here we collected fresh blood samples from 121 healthy donors, most with African ancestry, and performed exome sequencing, detailed RBC phenotyping, and parasite fitness assays. Over one third of healthy donors unknowingly carried alleles for G6PD deficiency or hemoglobinopathies, which were associated with characteristic RBC phenotypes. Among non-carriers alone, variation in RBC hydration, membrane deformability, and volume was strongly associated with P. falciparum growth rate. Common genetic variants in PIEZO1, SPTA1/SPTB, and several P. falciparum invasion receptors were also associated with parasite growth rate. Interestingly, we observed little or negative evidence for divergent selection on non-pathogenic RBC variation between Africans and Europeans. These findings suggest a model in which globally widespread variation in a moderate number of genes and phenotypes modulates P. falciparum fitness in RBCs.