Streptococcus mutans
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PROVIDER: PRJNA114941 | ENA |
REPOSITORIES: ENA
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Project description:Global transcriptional analysis of acid-inducible genes in Streptococcus mutans: multiple two-component systems involved in acid adaptation pH is a major environmental factor that regulates gene expression in many bacteria. Streptococcus mutans in dental biofilms is regularly exposed to cycles of acidic pH during the ingestion of fermentable dietary carbohydrates. The ability of S. mutans to tolerate low pH is crucial for its virulence and the pathogenesis in dental caries. To better understand its acid tolerance mechanisms, we used DNA microarray to perform genome-wide transcriptional analysis of S. mutans in response to acidic pH. The results showed that adaptation of S. mutans to pH 5.5 for 2 hrs induced differential expression of nearly 14% of genes in the genome, including 169 up-regulated genes and 108 down-regulated genes, largely categorized into six groups. Especially, we found that the genes encoding multiple two-component systems, including CiaHR, LevSR, LiaSR, ScnKR, HK/RR07 and ComDE, were up-regulated during acid adaptation. These findings were further confirmed by real time qRT-PCR and phenotypic assays of the gene deletion mutants. The results support that the multiple two-component systems are required for S. mutans to orchestrate its signal transduction networks for optimal adaptation to acidic pH.
2009-03-06 | GSE15125 | GEO
Project description:Global transcriptional analysis of acid-inducible genes in Streptococcus mutans: multiple two-component systems involved in acid adaptation pH is a major environmental factor that regulates gene expression in many bacteria. Streptococcus mutans in dental biofilms is regularly exposed to cycles of acidic pH during the ingestion of fermentable dietary carbohydrates. The ability of S. mutans to tolerate low pH is crucial for its virulence and the pathogenesis in dental caries. To better understand its acid tolerance mechanisms, we used DNA microarray to perform genome-wide transcriptional analysis of S. mutans in response to acidic pH. The results showed that adaptation of S. mutans to pH 5.5 for 2 hrs induced differential expression of nearly 14% of genes in the genome, including 169 up-regulated genes and 108 down-regulated genes, largely categorized into six groups. Especially, we found that the genes encoding multiple two-component systems, including CiaHR, LevSR, LiaSR, ScnKR, HK/RR07 and ComDE, were up-regulated during acid adaptation. These findings were further confirmed by real time qRT-PCR and phenotypic assays of the gene deletion mutants. The results support that the multiple two-component systems are required for S. mutans to orchestrate its signal transduction networks for optimal adaptation to acidic pH. Total RNAs were isolated from S. mutans UA159 cells (0.6 at OD600) grown in a TYG broth (3% tryptone, 0.3% yeast extract and 20 mM glucose) at either pH 5.5 or pH 7.5. The RNAs were treated with RNase-free DNase 1 and purified by Qiagen RNeasy mini columns. The purified RNAs were used to generate cDNA probes by an indirect labeling method based on the protocol from TIGR. The cDNAs were coupled with AlexaFluor 555 or AlexaFluor 647 (Invitrogen). The labeled cDNA probes from three different cultures of UA159 were hybridized to the S. mutans microarray slides obtained from PFGRC (http://pfgrc.tigr.org). Array hybridization was conducted using a protocol from the PFGRC with minor modification. After hybridization, washes and dried, the array slides were scanned by ScanArray 5000XL Reader (Perkin Elmer, Boston, MA). After the array slides were scanned, the resulting images were loaded into TIGR Spotfinder software (http://www.tigr.org/software/) and overlaid. A spot grid was created according to TIGR specifications and manually adjusted to fit all spots within the grid, and the intensity values of each spot were determined. Signal intensities of individual channels from an array slide were averaged and normalized using an array data analysis software (MIDAS) by using LOWESS and iterative log mean centering with default settings, followed by in-slide replicate analysis. A t-test was used to determine the consistency of ratios across replicate hybridizations. Only genes whose ratios were ⥠2-fold changes (either increase or decrease) with 99% confidence interval (P ⤠0.01) were considered statistically significant.
2010-05-22 | E-GEOD-15125 | biostudies-arrayexpress
Project description:Dental caries are closely associated with the virulence of Streptococcus mutans. The virulence expression of S. mutans is linked to its stress adaptation to the changes in the oral environment. In this work we used whole-genome microarrays to profile the dynamic transcriptomic responses of S. mutans during physiological heat stress. In addition, we evaluated the phenotypic changes, including initial biofilm formation, acid production and ATP turnover of S. mutans during heat stress. There were distinct patterns observed in the way that S. mutans responded to heat stress that included 66 transcription factors for the expression of functional genes being differentially expressed. Especially, response regulators of two component systems (TCSs), the repressors of heat shock proteins and regulators involved in sugar transporting and metabolism co-ordinated to enhance the cell’s survival and energy generation against heat stress in S. mutans.
2014-10-01 | GSE59302 | GEO
Project description:Bacteria can detect, transmit and react to signals from the outside world by using two-component systems an serine-threonine kinases and phosphatases. Streptococcus mutans contains one serine-threonine kinase, encoded by pknB. A gene encoding a serine-threonine phosphatase, pppL, is located downstream of pknB. In this study, the phenotypes of single mutants in pknB and pppL and a pknBpppL double mutant were characterized. All mutants exhibited a reduction in genetic transformability and biofilm formation, showed abnormal cell shapes, grew slower than the wild type strain in several complex media and had lost acid tolerance. The mutants had reduced cariogenic capacity, but no defects in colonization in a rat caries model. Whole genome transcriptome analysis revealed that pknB mutant showed reduced expression of genes involved in bacteriocin production and genetic competence. Among the genes that were diferentially regulated in the pknB mutant, severeal were likely to be involved in cell wall metabolism. One such gene, SMU.2146c and two genes encoding bacteriocins, were showed to be also down-regulated in vicK, which encodes a sensor kinase involved in response to oxidative stress. Collectively, the results suggest that PknB can modulate the activity of the two-component signal transduction systems vicKR and comDE. Real-time PCR showed that the genes down-regulated in the pknB mutant were up-regulated in the pppL mutant, indicating that PppL served to counteract PknB. Wild-type strain UA159 and a pknB mutant derivative were grown planktonically until OD600nm was 0.3. RNA was extracted, converted to cDNA, labelled and hybridized on S. mutans microarrays. The experiment was done in triplicate including one dye swap.
2010-05-06 | E-GEOD-18355 | biostudies-arrayexpress
Project description:Dental caries is closely associated with the virulence of Streptococcus mutans (S. mutans). The stress adaptation of S. mutans in the fluctuating oral environment is critical to the virulence expression of this bacterium. Here we used whole-genome microarrays to profile the dynamic transcriptomic responses of S. mutans during physiological heat stress. We also evaluated the phenotypic changes, including initial biofilm formation, acid production and ATP turnover of S. mutans during heat stress. We found that S. mutans responded to heat stress in a distinct pattern, which featured a differential transcription of 885 genes in total and 114 “core” transcripts throughout the six post-exposure time points investigated (5 min, 10 min, 15 min, 30 min, 45 min and 60 min). Further gene ontology analysis showed that transcriptional changes of genes involved in transcriptional regulation and cellular homeostasis were critical for heat stress responses of S. mutans. In addition to those highly conserved heat-shock proteins, we observed differential expression of multiple transcriptional regulators. The expression of genes involved in sugar transport, soluble glucans biosynthesis (gtfC) and binding (gbpC) were upregulated, whereas genes involved in ABC transporters, insoluble glucans biosynthesis (gtfB) and binding (gbpB), which are critical for biofilm skeleton, were either down-regulated or unchanged. These results together with enhanced glycolytic activity, attenuated sucrose-dependent initial attachment and impaired biofilm architecture indicate metabolic adaptations by this bacterium to compensate the extra energy demand for a better fitness under adverse conditions. We used time series microarrays to detect the dynamic changes of S. mutans under heat stress. Mid-logarithmic phase cell cultures of S. mutans (OD600nm = 0.5) were incubated at 42℃ for 5 min, 10 min, 15 min, 30 min, 45 min and 60 min, which were compared with the S. mutans cultured at 37℃. We applied three biological replicates at each time point.
2014-10-01 | E-GEOD-59302 | biostudies-arrayexpress
Project description:Bacteria can detect, transmit and react to signals from the outside world by using two-component systems an serine-threonine kinases and phosphatases. Streptococcus mutans contains one serine-threonine kinase, encoded by pknB. A gene encoding a serine-threonine phosphatase, pppL, is located downstream of pknB. In this study, the phenotypes of single mutants in pknB and pppL and a pknBpppL double mutant were characterized. All mutants exhibited a reduction in genetic transformability and biofilm formation, showed abnormal cell shapes, grew slower than the wild type strain in several complex media and had lost acid tolerance. The mutants had reduced cariogenic capacity, but no defects in colonization in a rat caries model. Whole genome transcriptome analysis revealed that pknB mutant showed reduced expression of genes involved in bacteriocin production and genetic competence. Among the genes that were diferentially regulated in the pknB mutant, severeal were likely to be involved in cell wall metabolism. One such gene, SMU.2146c and two genes encoding bacteriocins, were showed to be also down-regulated in vicK, which encodes a sensor kinase involved in response to oxidative stress. Collectively, the results suggest that PknB can modulate the activity of the two-component signal transduction systems vicKR and comDE. Real-time PCR showed that the genes down-regulated in the pknB mutant were up-regulated in the pppL mutant, indicating that PppL served to counteract PknB.
2010-03-05 | GSE18355 | GEO
Project description:Transcriptional Profiling of Streptococcus mutans UA159 Grown in Continuous Culture using TV Media Supplemented With 10 mM vs 100 mM Glucose. The genetic and phenotypic responses of Streptococcus mutans, an organism known to be strongly associated with the development of dental caries, to changes in carbohydrate availability were investigated. S. mutans UA159 or a derivative of UA159 lacking ManL, which is the EIIAB component (EIIABMan) of a mannose/glucose permease of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) and a dominant effector of catabolite repression, were grown in continuous culture to steady-state in conditions of excess (100 mM) or limiting (10 mM) glucose. Microarrays using RNA from S. mutans UA159 revealed that 174 genes were differentially expressed in response to changes in carbohydrate availability (P < 0.001). Glucose-limited cells possessed higher PTS activity, could acidify the environment more rapidly and to a greater extent, and produced more ManL protein than cultures grown with excess glucose. Loss of ManL adversely affected carbohydrate transport and acid tolerance. Comp arison of the HPr protein in S. mutans UA159 and the manL deletion strain indicated that the differences in behaviors of the strains were not due to major differences in HPr pools or HPr phosphorylation status. Therefore, carbohydrate availability alone can dramatically influence the expression of physiologic and biochemical pathways that contribute directly to the virulence of S. mutans, and ManL has a profound influence on this behavior. Two-condition experiment, growth in 10 mM vs 100 mM glucose. Biological replicates: 3 per condition, independently grown and harvested. One replicate per array
2013-12-03 | E-GEOD-51362 | biostudies-arrayexpress
Project description:Despite the widespread use of antiseptics such as chlorhexidine digluconate (CHX) in dental practice and oral care, the risks of potential resistance toward antiseptics in oral bacteria have only been highlighted very recently. Since the molecular mechanisms behind antiseptic resistance or adaptation are not entirely clear yet and the bacterial stress response has not been investigated systematically so far, the aim of the present study was to investigate the transcriptomic stress response in Streptococcus mutans after treatment with CHX using RNA sequencing. Planktonic cultures of stationary phase S. mutans were treated with a sublethal dose of CHX (125 µg/mL) for 5 min. After treatment, RNA was extracted, and RNA sequencing was performed on the Illumina NextSeq 500. Differential expressed genes were analyzed and validated by qRT-PCR. The analysis of the differential gene expression following pathway analysis revealed a considerable number of genes and pathways significantly regulated in S. mutans after sublethal treatment with CHX. In summary, expression of 423 genes was up-regulated and 295 genes down-regulated after CHX treatment. Analysis of differentially expressed genes and significantly regulated pathways showed regulation of genes involved in purine nucleotide synthesis, biofilm formation, transport systems and stress responses. In conclusion, these results show an overview of the transcriptomic stress response in S. mutans upon exposure to CHX and give an insight in potential mechanisms that may result in development of resistances.
2022-03-30 | GSE197633 | GEO
Project description:In the human pathogen Streptococcus mutans, the canonical peptide-based quorum sensing system is an inducible DNA repair system that is pivotal for bacterial survival. Previous work showed that the CSP signaling peptide is a stress-signaling alarmone that controls different stress-induced phenotypes. In this study, we exposed S. mutans to the CSP pheromone to mimic DNA damage conditions. Transcriptome analysis was then performed to evaluate the differential gene expression between the normal stationary phase cells and the CSP-induced stationary phase cells. The data obtained contribute to the understanding of the CSP-induced phenotypes in S. mutans.
2024-04-28 | GSE264670 | GEO
Project description:An essential two-component system WalRK is required for biofilm formation of streptococcus mutans mainly through transcriptional regulations.
2021-09-30 | PXD028893 |