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Functional Analysis of a Fibronectin Binding Protein of Streptococcus parasanguinis FW213

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Abstract

Streptococcus parasanguinis is a primary colonizer of dental plaque and an opportunistic pathogen for subacute endocarditis. A putative fibronectin binding protein (Spaf_1409) that lacks both an N-terminal signal peptide and a C-terminal cell wall-anchoring motif was identified from the S. parasanguinis FW213 genome. Spaf_1409 was abundantly present in the cytoplasm and also was found in the cell wall preparation and culture supernatant. By using an isogenic mutant strain, MPH4, Spaf_1409 was found to mediate the binding of S. parasanguinis FW213 to fibronectin. Inactivation of Spaf_1409 did not significantly alter the mass of static biofilm, but reduced the resistance of S. parasanguinis against the shearing force in a flow cell biofilm system, resulting in scattered biofilm. The mortality in Galleria mellonella larvae infected with MPH4 was higher than in those infected with wild-type S. parasanguinis. However, fewer viable bacterial cells were recovered from larvae infected with MPH4, compared to those infected with wild-type S. parasanguinis, up to 42 h post infection, suggesting that the infection by MPH4, but not the growth, was responsible for the elevated mortality. The phagocytic analysis using flow cytometry indicated that Spaf_1409 participates in the recognition of S. parasanguinis FW213 by RAW264.7 macrophages, suggesting that inactivation of Spaf_1409 intensified the immune responses in larvae, leading to larval death. Taken together, the data indicate that Spaf_1409 plays different roles in the development of dental biofilm and in systemic infections.

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Acknowledgements

This work was supported by the Chang Gung Memorial Hospital of Taiwan, Grant CMRPD1J0041-3 and 1J0301 to Y. M. Chen. We thank R. Faustoferri for review of the manuscript.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, College of Medicine, Chang Gung University, 259 Wen-Hwa 1st Road, Guishan Dist., Taoyuan City 333, Taiwan

    Yi-Ywan M. Chen

  2. Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan

    Yi-Ywan M. Chen, Pei-Shan Lu & Pei-Hua Tsai

  3. Molecular Infectious Disease Research Center, Chang Gung Memorial Hospital, Linkou, Taiwan

    Yi-Ywan M. Chen & Cheng-Hsun Chiu

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Conceived and designed the experiments: YMC and PL. Performed the experiments: PL and PT. Analyzed the data: YMC, PL, and PT. Contributed reagents/materials/analysis tools: CC. Wrote the paper: YMC and PL.

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Correspondence to Yi-Ywan M. Chen.

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Chen, YY.M., Lu, PS., Tsai, PH. et al. Functional Analysis of a Fibronectin Binding Protein of Streptococcus parasanguinis FW213. Curr Microbiol 77, 3430–3440 (2020). https://doi.org/10.1007/s00284-020-02152-7

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