Actinomyces spp. gene expression in root caries lesions

Background: The studies of the distribution of Actinomyces spp. on carious and non-carious root surfaces have not been able to confirm the association of these bacteria with root caries, although they were extensively implicated as a prime suspect in root caries. Objective: The aim of this study was to observe the gene expression of Actinomyces spp. in the microbiota of root surfaces with and without caries. Design: The oral biofilms from exposed sound root surface (SRS; n = 10) and active root caries (RC; n = 30) samples were collected. The total bacterial RNAwas extracted, and the mRNAwas isolated. Samples with low RNA concentration were pooled, yielding a final sample size of SRS = 10 and RC = 9. Complementary DNA (cDNA) libraries were prepared and sequenced on an Illumina (R) HiSeq 2500 system. Sequence reads were mapped to eight Actinomyces genomes. Count data were normalized using DESeq2 to analyse differential gene expression applying the Benjamini-Hochberg correction (false discovery rate [FDR] < 0.001). Results: Actinomyces spp. had similar numbers of reads (Mann-Whitney U-test; p > 0.05), except for Actinomyces OT178 (p = 0.001) and Actinomyces gerencseriae (p = 0.004), which had higher read counts in the SRS. Genes that code for stress proteins (clp, dnaK, and groEL), enzymes of glycolysis pathways (including enolase and phosphoenolpyruvate carboxykinase), adhesion (Type-2 fimbrial and collagen-binding protein), and cell growth (EF-Tu) were highly - but not differentially (p > 0.001) - expressed in both groups. Genes with the most significant upregulation in RC were those coding for hypothetical proteins and uracil DNA glycosylase (p = 2.61E-17). The gene with the most significant upregulation in SRS was a peptide ABC transporter substrate-binding protein (log2FC = -6.00, FDR = 2.37E-05). Conclusion: There were similar levels of Actinomyces gene expression in both sound and carious root biofilms. These bacteria can be commensal in root surface sites but may be cariogenic due to survival mechanisms that allow them to exist in acid environments and to metabolize sugars, saving energy.