Dental caries is a chronic infectious disease of multifactorial etiology that derives from the interplay among cariogenic bacteria on the dentition, the host diet, and other environmental exposures. Streptococcus mutans proliferates as a biofilm on the tooth surface, where it obtains nutrients and metabolizes fermentable dietary carbohydrates. The accumulation of lactic acid as a by-product of fermentation results in acidification of the plaque biofilm and demineralization of tooth enamel, marking the onset of decay. The ability of S. mutans to respond to environmental stresses presented by salivary flow, acid pH, oxidative stress, and changes in carbohydrate source and availability is essential for its survival and predominance in caries lesions. Importantly, S. mutans has evolved a network of regulators to integrate its cellular response to environmental change. Herein we describe the latest insights into global gene regulation in S. mutans, including mechanisms of signal transduction, carbon catabolite repression, and quorum-sensing. An improved understanding of these regulatory networks can provide a basis for novel therapeutic applications aimed at treating and/or preventing caries.

Streptococcus mutans is a commensal member of the healthy plaque biofilm and the primary causative agent of dental caries. The present study is an investigation of SloR, a 25-kDa metalloregulatory protein that modulates genes responsible for S. mutans-induced cariogenesis. Previous studies of SloR homologues in other bacterial pathogens have identified three domains critical to repressor functionality: an N-terminal DNA-binding domain, a central dimerization domain, and a C-terminal FeoA (previously SH3-like) domain. We used site-directed mutagenesis to identify critical amino acid residues within each of these domains of the SloR protein. Select residues were targeted for mutagenesis, and nonconservative amino acid substitutions were introduced by overlap extension PCR. Furthermore, three C-terminally truncated SloR variants were generated using conventional PCR. The repressor functionality and DNA-binding ability of each variant was assessed using CAT reporter gene assays, real-time semiquantitative reverse transcriptase (qRT)-PCR, and electrophoretic mobility shift assays. We identified 12 residues within SloR that cause significant derepression of sloABC promoter activity (P < 0.05) compared to the results for wild-type SloR. Derepression was particularly noteworthy in metal ion-binding site 1 mutants, consistent with the site's importance in gene repression by SloR. In addition, a hyperactive SloR(E169A/Q170A) mutant was identified as having significantly heightened repression of sloABC promoter activity, and experiments with C-terminal deletion mutants support involvement of the FeoA domain in SloR-mediated gene repression. Given these results, we describe the functional domains of the S. mutans SloR protein and propose that the hyperactive mutant could serve as a target for rational drug design aimed at repressing SloR-mediated virulence gene expression.A s a resident member of the healthy plaque biofilm and the principal contributor to dental caries formation in humans, Streptococcus mutans is well adapted to survive in the feast-andfamine conditions of the oral cavity (1). While there are many factors correlated with S. mutans pathogenesis, few play a more crucial role than metal ion homeostasis. Divalent metal ions, such as Fe 2ϩ and Mn 2ϩ , serve as important enzyme cofactors and as stabilizing components of proteins in a variety of key metabolic and cariogenic pathways (2, 3). In S. mutans, manganese has been shown to stimulate the conversion of dietary carbohydrates to acid and intracellular polysaccharide storage polymers, both of which are significant contributors to the caries-forming process (4). Moreover, the addition of manganese to drinking water significantly increased caries incidence in rats singly infected with S. mutans (4). Consequently, the sequestering and regulation of metal ions in the highly variable microenvironment of the oral cavity are paramount for the persistence and cariogenicity of S. mutans.Genome-wide sequence analysis of the S. mutans UA159 wildtype progenitor strain rev...