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The dental caries pathogenStreptococcus mutans is continually exposed to several types of stress in the oral biofilm environment. Oxidative stress generated by reactive oxygen species has a major impact on the establishment, persistence, and virulence ofS. mutans . Here, we combined fluorescent reporter-promoter fusions with single-cell imaging to study the effects of reactive oxygen species on activation of genetic competence inS. mutans . Exposure to paraquat, which generates superoxide anion, produced a qualitatively different effect on activation of expression of the gene for the master competence regulator, ComX, than did treatment with hydrogen peroxide (H2 O2 ), which can yield hydroxyl radical. Paraquat suppressed peptide-mediated induction ofcomX in a progressive and cumulative fashion, whereas the response to H2 O2 displayed a strong threshold behavior. Low concentrations of H2 O2 had little effect on induction ofcomX or the bacteriocin genecipB , but expression of these genes declined sharply if extracellular H2 O2 exceeded a threshold concentration. These effects were not due to decreased reporter gene fluorescence. Two different threshold concentrations were observed in the response to H2 O2 , depending on the gene promoter that was analyzed and the pathway by which the competence regulon was stimulated. The results show that paraquat and H2 O2 affect theS. mutans competence signaling pathway differently, and that some portions of the competence signaling pathway are more sensitive to oxidative stress than others.IMPORTANCE Streptococcus mutans inhabits the oral biofilm, where it plays an important role in the development of dental caries. Environmental stresses such as oxidative stress influence the growth ofS. mutans and its important virulence-associated behaviors, such as genetic competence.S. mutans competence development is a complex behavior that involves two different signaling peptides and can exhibit cell-to-cell heterogeneity. Although oxidative stress is known to influenceS. mutans competence, it is not understood how oxidative stress interacts with the peptide signaling or affects heterogeneity. In this study, we used fluorescent reporters to probe the effect of reactive oxygen species on competence signaling at the single-cell level. Our data show that different reactive oxygen species have different effects onS. mutans competence, and that some portions of the signaling pathway are more acutely sensitive to oxidative stress than others.

Oxidative Stressors Modify the Response of Streptococcus mutans to Its Competence Signal Peptides