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Highly-conserved regulatory activity of the ANR family in the virulence of diarrheagenic bacteria through interaction with Master and Global regulators

ABSTRACT: To gain mechanistic insights into this phenomenon, we characterized the regulatory role of Rnr in the virulence of prototype EPEC strain E2348-69 by genetic, biochemical, and human organoid-based approaches. Accordingly, RNA-seq analysis revealed more than 500 genes differentially regulated by Rnr, including the type-3 secretion system (T3SS).

ORGANISM(S): Escherichia coli

PROVIDER: GSE225626 | GEO | 2023/05/03

REPOSITORIES: GEO

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Comparative genomics provides insight into the diversity of the attaching and effacing Escherichia coli (AEEC) virulence plasmids

Project description:Global transcriptional analysis of the EPEC prototype strain E2348/69 and its plasmid mutants containing a deletion of perABC, or missing the entire EAF plasmid (strain JPN15).

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Transcriptional response induced by enteropathogenic Escherichia coli (EPEC) in mouse intestinal epithelial cells in vivo

Project description:In order to identify genes that were differentally regulated upon oral infection with EPEC, we isolated 8 days post-infection intestinal epithelial cells (IECs) from the small intestine of C57BL/6 neonate mice that were left untreated or orally infected with 5x104 WT EPEC E2348/69 or with 5x104 of the isogenic mutant ΔescV on their first day of life.

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Project description:Enterohemorrhagic Escherichia coli (EHEC) is a gram negative enteric bacterial pathogen that can cause hemorrhagic colitis and heamolytic uremic syndrome (HUS) in humans and is the cause of bloody diarrhoea and acute renal failure in children. We have studied the transcriptional response of a colon cell line (CaCo2) to infection by EHEC and another closely related enteric pathogen Enteropathogenic Escherichia coli (EPEC) and compared its response to a cervical cell line (Hela). We carried out microarray analysis on CaCo2 infected with EHEC O157H:7 EDL933 and EPEC E2348/69 at 4 hours of infection and analysis on Hela infected with EHEC also at 4 hours of infection

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Cell type dependent comparison and strain specific comparison

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RIL-seq of EPEC E2348/69 hfq-flag, in virulence activating and non-activating conditions.

Project description:RIL-seq experiment of EPEC hfq-flag mutant, in activating- conditions:growth on DMEM at 37°C to mid exponential growth phase (e.g., OD600=0.3). In these conditions EPEC strongly expresses its major virulence components, T3SS and BFP, mimicking infection. Non-activating conditions: overnight growth of static culture on LB medium at 37°C where virulence factors are not expressed. RIL-seq experiments are designed to reveal the interactions of sRNA and their targets.

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RNA-seq of EPEC E2348/69 in virulence activating and non-activating conditions. Comparison of wt and dhfq strains.

Project description:Compared the transcriptome of wild type EPEC with that of an isogenic Δhfq mutant. Comparing activating- conditions:growth on DMEM at 37°C to mid exponential growth phase (e.g., OD600=0.3). In these conditions EPEC strongly expresses its major virulence components, T3SS and BFP, mimicking infection. Non-activating conditions: overnight growth of static culture on LB medium at 37°C where virulence factors are not expressed.

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RIL-seq of EPEC E2348/69 hfq-flag, in virulence activating and non-activating conditions.

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Metabolome and transcriptome-wide effects of the carbon storage regulator A in enteropathogenic Escherichia coli.

Project description:The carbon storage regulator A (CsrA) is a conserved swivel of a global regulatory system known to regulate central carbon pathways, biofilm formation, motility, and pathogenicity. The aim of this study was to characterize changes in major metabolic pathways induced by CsrA in the human enteropathogenic Escherichia coli (EPEC) strain E2348/69. The EPEC strain E2348/69 and a csrA deletion mutant were grown under virulence factor inducing conditions and characterized by a combined analysis of their metabolomes and transcriptomes. Of the 159 metabolites identified from untargeted GC/MS and LC/MS data, 70 were significantly (fold change ≥ 1.5; p-value ≤ 0.05) regulated between the knockout and the wildtype strain. A lack of csrA led to an upregulation of upper glycolysis and glycogen synthesis pathways, whereas lower glycolysis and the citric acid cycle were downregulated. Associated pathways from the citric acid cycle like aromatic amino acid and siderophore biosynthesis were also negatively influenced. The nucleoside salvage pathways were featured by an accumulation of nucleosides and nucleobases, and a downregulation of nucleotides. In addition, a pronounced downregulation of lyso-lipid metabolites was observed. A drastic change in the morphology in the form of vesicle-like structures of the csrA knockout strain was visible by electron microscopy, which is supposed to be a consequence of a strong upregulation of colanic acid synthesis. The findings expand the scope of pathways affected by the csrA regulon and emphasize its importance as a global regulator.

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