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The Oral Bacterium Fusobacterium nucleatum Binds Staphylococcus aureus and Alters Expression of the Staphylococcal Accessory Regulator sarA

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Abstract

Staphylococcus aureus, an opportunistic pathogen member of the nasal and skin microbiota, can also be found in human oral samples and has been linked to infectious diseases of the oral cavity. As the nasal and oral cavities are anatomically connected, it is currently unclear whether S. aureus can colonize the oral cavity and become part of the oral microbiota, or if its presence in the oral cavity is simply transient. To start addressing this question, we assessed S. aureus ability to directly bind selected members of the oral microbiota as well as its ability to integrate into a human-derived complex oral microbial community in vitro. Our data show that S. aureus forms aggregates with Fusobacterium nucleatum and Porphyromonas gingivalis and that it can incorporate into the human-derived in vitro oral community. Further analysis of the F. nucleatum-S. aureus interaction revealed that the outer-membrane adhesin RadD is partially involved in aggregate formation and that the RadD-mediated interaction leads to an increase in expression of the staphylococcal global regulator gene sarA. Our findings lend support to the notion that S. aureus can become part of the complex microbiota of the human mouth, which could serve as a reservoir for S. aureus. Furthermore, direct interaction with key members of the oral microbiota could affect S. aureus pathogenicity contributing to the development of several S. aureus associated oral infections.

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Acknowledgements

We thank members of the Lux, Weibel, and Herzberg laboratories for discussion and/or critical reading of the manuscript. We thank Dr. Blaise Boles for giving us S. aureus strains SH1000 and USA300. We also thank Dr. Kelly Schwartz for helping us optimize the growth conditions for S. aureus biofilm.

Funding

The study was supported by a grant from the National Institutes of Health: NIDCR R01 DE021108 (R. L.) with a research supplement (B. L.) and by a grant from the National Science Foundation DMR-1121288 (D. W.). The funding agency had no role in study design, data collection, and interpretation or the decision to submit the work for publication.

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Author notes

  1. Bruno P. Lima & Gerrit W. Vreeman

    Present address: Department of Diagnostic and Biological Sciences, School of Dentistry, Universit of Minnesota, Minneapolis, MN, USA

  2. Linda I. Hu

    Present address: Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA

  3. Bruno P. Lima and Linda I. Hu contributed equally to this work.

Authors and Affiliations

  1. Division of Constitutive and Regenerative Sciences, School of Dentistry, University of California, Los Angeles, CA, USA

    Bruno P. Lima & Renate Lux

  2. Department of Biochemistry, University of Wisconsin-Madison, 440 Henry Mall, Madison, WI, USA

    Linda I. Hu & Douglas B. Weibel

  3. Department of Chemistry, University of Wisconsin-Madison, 440 Henry Mall, Madison, WI, USA

    Douglas B. Weibel

  4. Department of Biomedical Engineering, University of Wisconsin-Madison, 440 Henry Mall, Madison, WI, USA

    Douglas B. Weibel

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Correspondence to Renate Lux.

Electronic Supplementary Material

Supplemental Fig. 1)

Biofilm formation of S. aureus SH1000 (Sa) cells, TBS66 media, compared to media alone or empty well. The plate was incubated anaerobically at 37 °C for 16 h. Crystal violet retention was used as an indicator of biomass. Data represent means and standard deviation of three independent replicates. (JPG 11 kb)

Supplemental Fig. 2)

aS. aureus SH1000 was grown shaking with aeration in TSB66 containing 0–0.3% maltose w/v in a 96 well plate for 24 h. Biofilm was quantified by crystal violet staining of attached cells (left Y-axis). Growth of the planktonic cells was quantified by transferring the cultures into a clean 96 well plate and measuring the absorbance (right Y-axis). The values are averages of four wells and standard deviations. bS. aureus SH1000 was grown at 37 °C, shaking with aeration in TSB66 containing 0–0.3% maltose w/v for 10 h. OD600 was measured every 50 min. Data represent means and standard deviation of 3 biological replicates. (JPG 42 kb)

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Lima, B.P., Hu, L.I., Vreeman, G.W. et al. The Oral Bacterium Fusobacterium nucleatum Binds Staphylococcus aureus and Alters Expression of the Staphylococcal Accessory Regulator sarA. Microb Ecol 78, 336–347 (2019). https://doi.org/10.1007/s00248-018-1291-0

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