Novel Two-Component System ofStreptococcus sanguinisAffecting Functions Associated with Viability in Saliva and Biofilm Formation
ABSTRACTStreptococcus sanguinisis a pioneer species of teeth and a common opportunistic pathogen of infective endocarditis. In this study, we identified a two-component system,S. sanguinisSptRS (SptRSSs), affectingS. sanguinissurvival in saliva and biofilm formation. Isogenic mutants ofsptRSs(SKsptR) andsptSSs(SKsptS) showed reduced cell counts inex vivoassays of viability in saliva compared to those of parent strain SK36 and complemented mutants. Reduced counts of the mutants in saliva were associated with reduced growth rates in nutrient-poor medium (RPMI) and increased susceptibility to the deposition of C3b and the membrane attach complex (MAC) of the complement system, a defense component of saliva and serum. Conversely,sptRSsandsptSSsmutants showed increased biofilm formation associated with higher levels of production of H2O2and extracellular DNA. Reverse transcription-quantitative PCR (RT-qPCR) comparisons of strains indicated a global role of SptRSSsin repressing genes for H2O2production (2.5- to 15-fold upregulation ofspxB,spxR,vicR,tpk, andackAinsptRSsandsptSSsmutants), biofilm formation, and/or evasion of host immunity (2.1- to 11.4-fold upregulation ofsrtA,pcsB,cwdP,iga, andnt5e). Compatible with the homology of SptRSswith AraC-type regulators, duplicate to multiple conserved repeats were identified in 1,000-bp regulatory regions of downstream genes, suggesting that SptRSsregulates transcription by DNA looping. Significant transcriptional changes in the regulatory genesvicR,spxR,comE,comX, andmecAin thesptRSsandsptSSsmutants further indicated that SptRSSsis part of a regulatory network that coordinates cell wall homeostasis, H2O2production, and competence. This study reveals that SptRSSsis involved in the regulation of crucial functions forS. sanguinispersistence in the oral cavity.