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Research Article

Uncovering Roles of Streptococcus gordonii SrtA-Processed Proteins in the Biofilm Lifestyle

Brittany L. Nairn, Grace T. Lee, Ashwani K. Chumber, Patrick R. Steck, Mahmoud O. Mire, Bruno P. Lima, Mark C. Herzberg
Michael J. Federle, Editor
Brittany L. Nairn
bDepartment of Biological Sciences, Bethel University, St. Paul, Minnesota, USA
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Grace T. Lee
aDepartment of Biological and Diagnostic Sciences, University of Minnesota, Minneapolis, Minnesota, USA
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Ashwani K. Chumber
bDepartment of Biological Sciences, Bethel University, St. Paul, Minnesota, USA
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Patrick R. Steck
aDepartment of Biological and Diagnostic Sciences, University of Minnesota, Minneapolis, Minnesota, USA
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Mahmoud O. Mire
aDepartment of Biological and Diagnostic Sciences, University of Minnesota, Minneapolis, Minnesota, USA
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Bruno P. Lima
aDepartment of Biological and Diagnostic Sciences, University of Minnesota, Minneapolis, Minnesota, USA
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Mark C. Herzberg
aDepartment of Biological and Diagnostic Sciences, University of Minnesota, Minneapolis, Minnesota, USA
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Michael J. Federle
University of Illinois at Chicago
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DOI: 10.1128/JB.00544-20
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ABSTRACT

Streptococcus gordonii is a commensal oral organism. Harmless in the oral cavity, S. gordonii is an opportunistic pathogen. S. gordonii adheres to body surfaces using surface adhesive proteins (adhesins), which are critical to subsequent formation of biofilm communities. As in most Gram-positive bacteria, S. gordonii surface proteins containing the C-terminal LPXTG motif cleavage sequence are processed by sortase A (SrtA) to become covalently attached to the cell wall. To characterize the functional diversity and redundancy in the family of SrtA-processed proteins, an S. gordonii DL1 markerless deletion mutant library was constructed of each of the 26 putative SrtA-processed proteins. Each library member was evaluated for growth in rich medium, biofilm formation on plastic, saliva and salivary fractions, cell surface hydrophobicity (CSH), hemagglutination, and integration into an ex vivo plaque biofilm community. Library members were compared to the non-SrtA-processed adhesins AbpA and AbpB. While no major growth differences in rich medium were observed, many S. gordonii LPXTG/A proteins impacted biofilm formation on one or more of the substrates. Several mutants showed significant differences in hemagglutination, hydrophobicity, or fitness in the ex vivo plaque model. From the identification of redundant and unique functions in these in vitro and ex vivo systems, functional stratification among the LPXTG/A proteins is apparent.

IMPORTANCE S. gordonii interactions with its environment depend on the complement of cell wall proteins. A subset of these cell wall proteins requires processing by the enzyme sortase A (SrtA). The identification of SrtA-processed proteins and their functional characterization will help the community to better understand how S. gordonii engages with its surroundings, including other microbes, integrates into the plaque community, adheres to the tooth surface, and hematogenously disseminates to cause blood-borne infections. This study identified 26 putative SrtA-processed proteins through creation of a markerless deletion mutant library. The library was subject to functional screens that were chosen to better understand key aspects of S. gordonii physiology and pathogenesis.

FOOTNOTES

    • Received 30 September 2020.
    • Accepted 20 October 2020.
    • Accepted manuscript posted online 26 October 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

All Rights Reserved.

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Uncovering Roles of Streptococcus gordonii SrtA-Processed Proteins in the Biofilm Lifestyle
Brittany L. Nairn, Grace T. Lee, Ashwani K. Chumber, Patrick R. Steck, Mahmoud O. Mire, Bruno P. Lima, Mark C. Herzberg
Journal of Bacteriology Dec 2020, 203 (2) e00544-20; DOI: 10.1128/JB.00544-20

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Uncovering Roles of Streptococcus gordonii SrtA-Processed Proteins in the Biofilm Lifestyle
Brittany L. Nairn, Grace T. Lee, Ashwani K. Chumber, Patrick R. Steck, Mahmoud O. Mire, Bruno P. Lima, Mark C. Herzberg
Journal of Bacteriology Dec 2020, 203 (2) e00544-20; DOI: 10.1128/JB.00544-20
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KEYWORDS

streptococcus
biofilms
cell wall
oral microbiology
sortase A

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