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Molecular Pathogenesis | Spotlight

Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence

Norhan Alhajjar, Anushila Chatterjee, Brady L. Spencer, Lindsey R. Burcham, Julia L. E. Willett, Gary M. Dunny, Breck A. Duerkop, Kelly S. Doran
Nancy E. Freitag, Editor
Norhan Alhajjar
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Anushila Chatterjee
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Brady L. Spencer
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Lindsey R. Burcham
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Julia L. E. Willett
bDepartment of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, Minnesota, USA
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Gary M. Dunny
bDepartment of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, Minnesota, USA
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Breck A. Duerkop
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Kelly S. Doran
aDepartment of Immunology and Microbiology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, Colorado, USA
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Nancy E. Freitag
University of Illinois at Chicago
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DOI: 10.1128/IAI.00270-20
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ABSTRACT

Enterococcus faecalis is a Gram-positive commensal bacterium native to the gastrointestinal tract and an opportunistic pathogen of increasing clinical concern. E. faecalis also colonizes the female reproductive tract, and reports suggest vaginal colonization increases following antibiotic treatment or in patients with aerobic vaginitis. Currently, little is known about specific factors that promote E. faecalis vaginal colonization and subsequent infection. We modified an established mouse vaginal colonization model to explore E. faecalis vaginal carriage and demonstrate that both vancomycin-resistant and -sensitive strains colonize the murine vaginal tract. Following vaginal colonization, we observed E. faecalis in vaginal, cervical, and uterine tissue. A mutant lacking endocarditis- and biofilm-associated pili (Ebp) exhibited a decreased ability to associate with human vaginal and cervical cells in vitro but did not contribute to colonization in vivo. Thus, we screened a low-complexity transposon (Tn) mutant library to identify novel genes important for E. faecalis colonization and persistence in the vaginal tract. This screen revealed 383 mutants that were underrepresented during vaginal colonization at 1, 5, and 8 days postinoculation compared to growth in culture medium. We confirmed that mutants deficient in ethanolamine catabolism or in the type VII secretion system were attenuated in persisting during vaginal colonization. These results reveal the complex nature of vaginal colonization and suggest that multiple factors contribute to E. faecalis persistence in the reproductive tract.

FOOTNOTES

    • Received 6 May 2020.
    • Returned for modification 9 June 2020.
    • Accepted 26 July 2020.
    • Accepted manuscript posted online 10 August 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

All Rights Reserved.

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Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence
Norhan Alhajjar, Anushila Chatterjee, Brady L. Spencer, Lindsey R. Burcham, Julia L. E. Willett, Gary M. Dunny, Breck A. Duerkop, Kelly S. Doran
Infection and Immunity Sep 2020, 88 (10) e00270-20; DOI: 10.1128/IAI.00270-20

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Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence
Norhan Alhajjar, Anushila Chatterjee, Brady L. Spencer, Lindsey R. Burcham, Julia L. E. Willett, Gary M. Dunny, Breck A. Duerkop, Kelly S. Doran
Infection and Immunity Sep 2020, 88 (10) e00270-20; DOI: 10.1128/IAI.00270-20
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    • ABSTRACT
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KEYWORDS

E. faecalis
Enterococcus
T7SS
TnSeq
colonization
ethanolamine catabolism
vaginal tract

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