enteropathogenic e coli atcc 25922  (ATCC)

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: Growth curves represent OD600 measurements recorded every 30 minutes for 15 hours for EPEC O125ac:H6 WT, Δ qseB, Δ qseC, Δ qseB Δ qseC, Δ pmrA, Δ pmrB, Δ pmrA Δ pmrB, Δ pmrA Δ qseB, Δ pmrA Δ qseC, Δ pmrB Δ qseB, Δ pmrB Δ qseC (A) and Δ kdpD, Δ kdpE Δ kdpD Δ qseB, Δ kdpD Δ qseC, Δ kdpE Δ qseB and Δ kdpE Δ qseC (B). Error bars represent the Standard Deviation, n = 4

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques: Standard Deviation

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: Relative motility compared to the WT of eight replicates is depicted for EPEC O125ac:H6 Δ qseB, Δ qseC, Δ qseB Δ qseC, Δ pmrA, Δ pmrB, Δ pmrA Δ pmrB, Δ pmrA Δ qseB, Δ pmrA Δ qseC, Δ pmrB Δ qseB, Δ pmrB Δ qseC. EPEC O125ac:H6 WT motility is calculated as the relative motility of the WT per plate compared to the average motility of the WT in the experiment. Motility plates that exemplify the seen motility are depicted. * p-value <0.05; ** <0.01; *** <0.001; **** <0.0001 via one-way ANOVA with multiple comparisons, n = 8

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques:

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: Relative motility compared to the WT of eight replicates is depicted for EPEC O125ac:H6 Δ qseB, Δ qseC, Δ kdpD, Δ kdpE Δ kdpD Δ qseB, Δ kdpD Δ qseC, Δ kdpE Δ qseB and Δ kdpE Δ qseC . EPEC O125ac:H6 WT motility is calculated as the relative motility of the WT per plate compared to the average motility of the WT in the experiment. Motility plates that exemplify the seen motility are depicted. * p-value <0.05; ** <0.01; *** <0.001; **** <0.0001 via one-way ANOVA with multiple comparisons, n = 8

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques:

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: A. Chemical structure of AI-3 B. Relative motility compared to the non-treated control of EPEC O125ac:H6 in presence of 500 µM of epinephrine, dopamine, AI-3 and Fe 3+ . C. Comparison between the relative motility of of EPEC O125ac:H6 mutants compared to the WT in non-treated agar plates with the relative motility of EPEC O125ac:H6 mutants compared to the WT in agar plates containing 500 µM Fe 3+ . Motility plates of mutants that are affected by the presence of iron are depicted. * p-value <0.05; ** <0.01; *** <0.001; **** <0.0001 via one-way ANOVA with multiple comparisons,

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques: Control, Comparison

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: A. Differential expression according to the luciferase assay of the genes flhC, fliA, ler, recA and bla of all tested mutants. Significant differences are indicated by ++ > 2-fold; + 1-2-fold; . non-significant differences with WT; - 1-0.5-fold and - - < 0.5-fold B. Quantitative-PCR of genes flhC and fliA in EPEC O125ac:H6 Δ qseB, Δ qseC, and Δ pmrA Δ qseC to prove that luciferase assay was accurate. C. qPCR in EPEC O125ac:H6 Δ qseC and Δ pmrA Δ qseC to check qseB expression in these mutants. * p-value <0.05; ** <0.01; *** <0.001; **** <0.0001 via one-way ANOVA with multiple comparisons, n = 4

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques: Quantitative Proteomics, Luciferase, Real-time Polymerase Chain Reaction, Expressing

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: A. MIC data of colistin and polymyxin B (PBM) for EPEC O125ac:H6 WT, Δ qseB, Δ qseC, Δ qseB Δ qseC, Δ pmrA, Δ pmrB, Δ pmrA Δ pmrB, Δ pmrA Δ qseB, Δ pmrA Δ qseC, Δ pmrB Δ qseB, Δ pmrB Δ qseC., Δ kdpD, Δ kdpE Δ kdpD Δ qseB, Δ kdpD Δ qseC, Δ kdpE Δ qseB and Δ kdpE Δ qseC . B. Differential expression using qPCR of colistin and PBM resistance genes arnB and eptA in the mutants EPEC O125ac:H6 WT, Δ qseB, Δ qseC and Δ pmrA Δ qseC. C. In the promoter region of arn operon, both QseB-binding site (blue) and PmrA-binding site (orange) can be found. The start (ATG, red) of arnB gene is also shown. * p-value <0.05; ** <0.01; *** <0.001; **** <0.0001 via one-way ANOVA with multiple comparisons, n = 4

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques: Quantitative Proteomics, Binding Assay

Journal: bioRxiv

Article Title: Cross-talk between QseBC and PmrAB two-component systems is crucial for regulation of motility and colistin resistance in Enteropathogenic Escherichia coli

doi: 10.1101/2023.04.06.535820

Figure Lengend Snippet: A. In WT aEPEC, KdpD HK phosphorylates an unknown effector to activate flagella late-regulatory genes. PmrB activates its cognate RR PmrA, and the non-cognate QseB. QseC HK controls the level of activated QseB via de-phosphorylation and sequestration of activated QseB. Unphosphorylated QseB in EPEC WT is able to exert a basal repression of flhC, recA and ler genes. B. In Δ pmrA Δ qseC mutant, QseB is phosphorylated via PmrB, and cannot be deactivated by QseC. P∼QseB activates the expression of qseBC operon leading to QseB over-expression. In arn operon, PmrA usually blocks QseB binding-site, but in its absence phosphorylated QseB leads to activation of colistin resistance genes. Excess of phosphorylated QseB tightly represses flhC and recA genes. Created with Biorender.com

Article Snippet: Atypical Enteropathogenic E. coli strain O125ac:H6 (DSM8711) was acquired from the Leibniz Institute DSMZ collection located in Germany.

Techniques: De-Phosphorylation Assay, Mutagenesis, Expressing, Over Expression, Binding Assay, Activation Assay