Transcriptome of S.pyogenes induced by the signaling peptide SilCR
Ontology highlight
ABSTRACT:
ORGANISM(S): Streptococcus pyogenes
PROVIDER: GSE16961 | GEO | 2009/07/16
SECONDARY ACCESSION(S): PRJNA117725
REPOSITORIES: GEO
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Project description:Transcriptional regulation of S. pyogenes strain JS95, containing streptococcal invasion locus (sil) by the pheromone quorum-sensing peptide SilCR. Transcriptional profiling was conducted by comparing untreated JS95 culture with SilCR-treated cultures, using two concentrations of SilCR (10 or 0.05 µg/ml) and two incubation times (10 or 180 minutes). As a control, we used JS95-derived mutant deficient of the two-component system SilA-SilB that is necessary for SilCR signaling (ÎsilAB). Two-condition experiment: with vs. without SilCR of WT JS95 strain; without vs. with SilCR of ÎsilAB mutant. Two biological repeates were performed for experiment using WT JS95 treated with 10 µg/ml SilCR for 10 minutes.
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Project description:The objective of this study was to investigate which genes are important for Streptococcus pyogenes during intracellular survival in human macrophages. Streptococcus pyogenes is an important human pathogen, which has recently gained recognition as an intracellular microorganism during the course of severe invasive infections such as necrotizing fasciitis. Although the surface anchored M protein has been identified as a pivotal factor affecting phagosomal maturation and S. pyogenes survival within macrophages, the overall transcriptional profile required for the pathogen to adapt and persist intracellularly is yet unknown. To address this, gene expression profiles of S. pyogenes within human macrophages were determined and compared to those of extracellular bacteria using customized microarrays and real-time qRT-PCR. In order to model the early phase of infection involving adaptation to the intracellular compartment, samples were collected 2h post-infection and within 2 h post infection, the expression of 145 streptococcal genes was significantly altered in the intracellular environment. The majority of differentially regulated genes were associated with metabolic and energy-dependent processes. Key upregulated genes in early phase intracellular bacteria were ihk and irr, encoding a two-component gene regulatory system (TCS). We observed that an isogenic S. pyogenes mutant deficient in ihk/irr displayed significantly reduced bacterial counts as compared to wild-type bacteria following infection of macrophages. Comparison of gene expression of selected genes at 2h and 6h post-infection revealed a dramatic shift in response regulators over time with a down-regulation of ihk/irr genes concurrent with an upreguation of the well-studied covR/S two component regulator. In reinfection assays, intracellular bacteria from the 6h time point exhibited significantly greater survival within macrophages than did bacteria collected at the 2h time point. The findings illustrate how gene expression of S. pyogenes during the intracellular life cycle is fine-tuned by temporal expression of specific two-component systems. Five samples with three biological replicates are analysed. Each open reading frame in triplicate (three technical replicas per sample). Resulting in 6-9 data points per gene per condition. The extracellular bacteria are control samples and the internal control is the house-keeping gene gyrase.
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