Regulation of gtf Gene Expression in S mutans

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Streptococcus mutans is a causative agent of dental caries. It's ability to inflict damage is strongly linked to the production of long chain glucose polymers (glucans) derived from dietary sucrose which allow the bacteria possesses a family of genes that express enzymes called glucosyltransferases (GTF). There are three GTFs in S. mutans. They share 50 percent sequence identity and are encoded by the homologous gtfB, gtfC and gtfD genes. The gtfB and gtfC gene are found in direct repeat with a mere 198 base paries separation between coding sequences. Although mutations in either gtfB or gtfC produce a colonizing deficient phenotype, little is known about their regulation. Until recently, the only effector known to regulate these genes was sucrose, the substrate of the GTFs. Supplemented sucrose transiently induces a 3-fold increase in a gtfB transcriptional fusion. Now we have demonstrated that both gtfB and gtfC show coordinated growth phase- dependent expression. For both genes, expression peaks prior to exponential growth and falls over 100-fold by early stationary phase. In this proposal, we intend to further study this phenomenon including identifying critical cis and trans acting elements and through mutagenesis determine how these elements affect gtf gene expression. Since these gtf genes are involved in the colonization process, the genetic elements that are found here will need to be examined in terms of the bacteria's pathogenic mechanisms.