Abstract

Dental caries is a biofilm-dependent disease that largely relies on the ability of Streptococcus mutans to synthesize exopolysaccharides. Although the rnc gene is suggested to be involved in virulence mechanisms in many other bacteria, the information regarding it in S. mutans is very limited. Here, using deletion or overexpression mutant assay, we demonstrated that rnc in S. mutans significantly positively regulated exopolysaccharide synthesis and further altered biofilm formation. Meanwhile, the cariogenecity of S. mutans was decreased by deletion of rnc in a specific pathogen-free (SPF) rat model. Interestingly, analyzing the expression at mRNA level, we found the downstream vic locus was repressed by rnc in S. mutans. Using deep sequencing and bioinformatics analysis, for the first time, three putative microRNA-size small RNAs (msRNAs) targeting vicRKX were predicted in S. mutans. The expression levels of these msRNAs were negatively correlated with vicRKX but positively correlated with rnc, indicating rnc probably repressed vicRKX expression through msRNAs at the post-transcriptional level. In all, the results present that rnc has a potential role in the regulation of exopolysaccharide synthesis and can affect vicRKX expressions via post-transcriptional repression in S. mutans. This study provides an alternative avenue for further research aimed at preventing caries.

Highlights

  • Dental caries is one of the most prevalent and costly biofilm-dependent oral infectious diseases, affecting the majority of the world’s population (Selwitz et al, 2007)

  • Ligation constructs and the shuttle vector were separately introduced into S. mutans UA159 using competencesimulating peptide (CSP)-induced natural transformation, and transformants that were independently resistant to erythromycin and spectinomycin were selected for recombination into the chromosome using PCR, followed by nucleotide sequence analysis

  • We found that the expression of rnc was positively correlated with exopolysaccharide synthesis

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Summary

INTRODUCTION

Dental caries is one of the most prevalent and costly biofilm-dependent oral infectious diseases, affecting the majority of the world’s population (Selwitz et al, 2007). As exopolysaccharide is an important virulence factor, exopolysaccharide synthesis-related genes are optimal targets for the development of anti-caries compounds (Chau et al, 2015; Zhang et al, 2015) Among these genes, vicRKX, which are known to encode the VicRK signal transduction system (TCS), are crucial to the pathogenicity of S. mutans, especially for exopolysaccharide formation (Senadheera et al, 2005, 2007). Recent parallel studies have provided evidence for the high abundance of discrete microRNA-size small RNAs (msRNAs) in S. mutans and Escherichia coli (Lee and Hong, 2012; Kang et al, 2013) Overall, these findings suggest that msRNAs may participate in rnc-related regulation as post-transcriptional regulators in bacteria. This paper provides an alternative avenue for further caries prevention research

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