Supplementary Material for: Streptococcus mutans gene expression and functional profile in root caries: an RNA-seq study

The literature is still scarce on studies describing S. mutans global gene expression under clinical conditions such as those found on complex biofilms from sound root surfaces (SRS) and carious root surfaces (RC). This study aimed to investigate the S. mutans gene expression and functional profile within the metatranscriptome of biofilms from SRS and from RC in an attempt to identify enriched functional signatures potentially associated with healthy to disease transitioning process. Total RNA was extracted, and prepared libraries (SRS=10 and RC=9) were paired-end sequenced using the Illumina HiSeq2500. Read count assigned to each gene of the S. mutans UA159 strain were obtained. Differentially expressed genes (DEG) between SRS and RC were identified using the DESeq2 R package and weighted gene co-expression network analysis (WGCNA) was performed to explore and identify functional modules related to SRS and RC. We found seventeen DEG between SRS and RC samples, with three overexpressed in RC and related to membrane protein, alanyl-tRNA synthetase and GTP-binding protein with the remaining ones overexpressed in SRS samples and related to hypothetical protein, transposon integrase, histidine kinase, putative transporter, bacteriocin immunity protein, response regulator, 6-phospho-beta-galactosidase, purine metabolism and to transcriptional regulator. Key-functional modules were identified for SRS and RC conditions based on WCGNA, being 139 hub genes found on SRS key-module and 17 genes on RC key-module. Functional analysis of S. mutans within the metatranscriptome of biofilms from sound root and from carious root revealed a similar pattern of gene expression, and only a few genes have been differentially expressed between biofilms from sound root surfaces and from root carious lesions. However, S. mutans presented a greater functional abundance in the lesion samples. Some functional patterns related to sugar (starch, sucrose, fructose, mannose and lactose) and heterofermentative metabolisms, to cell-wall biosynthesis and to acid tolerance stress seem to be enriched on carious root surfaces conferring ecological advantages to S. mutans. Altogether, the present data suggest that a functional signature may be associated with carious root lesions.