Defense Date

2011

Document Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Microbiology & Immunology

First Advisor

Todd Kitten

Abstract

Streptococcus sanguinis is an important component of the dental plaque biofilm and is believed to play a beneficial role in the oral cavity. S. sanguinis is also a leading cause of infective endocarditis (IE), a potentially lethal infection of the cardiac valves. S. sanguinis possesses genetic competence, the ability to acquire exogenous DNA into its genome. In the well characterized system of S. pneumoniae, genetic competence requires a quorum sensing system encoded by the early comCDE genes, as well as late genes controlled by the alternate sigma factor, ComX. Previous studies in other streptococcal species have suggested functions for the com regulon apart from DNA uptake. Here we characterized functions of the S. sanguinis com regulon genes in genetic competence, IE virulence, and biofilm formation. Our findings indicated that the early regulatory genes and those under the control of ComX in S. sanguinis play similar roles in genetic competence as their orthologs in other competent streptococci; however the sequence and mechanism of processing of the quorum sensing signal, competence-stimulating peptide, CSP, were determined to be unique. Using a rabbit endocarditis model, we determined that the comCDE and comX genes were not required for virulence, bacteremia, or pathology under a variety of infection conditions. In contrast, examination of biofilms by microscopy and crystal violet staining indicated that S. sanguinis CSP enhanced biofilm formation in a comDE-dependent manner. Deletion of the early com gene SSA_0195 eliminated this effect, while expression of the gene from an inducible promoter increased biofilm formation in the absence of CSP. Deletion of the comX gene resulted in biofilms with increased staining, cell death, and profoundly altered structure. Treatment with DNase I reduced biofilm formation in a com-independent manner. Taken together, these results suggest that expression of SSA_0195 is both necessary and sufficient for CSP-dependent biofilm enhancement, and that the late gene activator, ComX, is required to maintain normal biofilm architecture. Our findings suggest the com regulon of S. sanguinis may be an important determinant of competitiveness in the mouth, where native CSP production may occur at levels sufficient to influence biofilm formation.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

August 2011

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