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High resolution definition of the Staphylococcus aureus WalKR regulon [ChIP-seq]

ABSTRACT: Here, we use a panel of defined walKR mutants and functional genomics including ChIP-seq to confirm a high-resolution consensus WalR DNA-binding motif and characterise a 61-gene direct regulon

ORGANISM(S): Staphylococcus aureus

PROVIDER: GSE212318 | GEO | 2022/09/01

REPOSITORIES: GEO

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High resolution definition of the Staphylococcus aureus WalKR regulon [RNA-seq]

Project description:Here, we use a panel of defined walKR mutants and functional genomics including ChIP-seq to confirm a high-resolution consensus WalR DNA-binding motif and characterise a 61-gene direct regulon

2022-09-01 | GSE212319 | GEO

Transcriptome analysis of the WalKR regulon in Staphylococcus aureus

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Transcriptome analysis of the WalKR regulon in Staphylococcus aureus

Project description:The WalKR regulon was studied by comparing genes expression in a strain producing a constitutively active WalR regulator (WalRc) versus the wild type strain carrying the empty expression vector. Hybridizations were performed with three independent biological replicates. Our data allowed to identify 165 genes differentially expressed in the walRc expressing strain with a P-value M-bM-^IM-$ 0.05 using Z-test and a threshold value of two-fold change in transcriptional levels. Among them, 108 genes are activated, and 57 genes are repressed. About 10% of these results were confirmed by quantitative real time PCR, revealing the high reliability of this overall study. Beyond the major effect of the WalKR system on genes expressing cell wall hydrolases, we have shown that it activates a large number of genes involved in virulence, and probably through its positive impact on the SaeRS two-component system, is a major activator of S. aureus virulence. Cells were grown in TSB rich medium added with 0.25 M-BM-5M CdCl2 to allow walRc expression until OD600nm of 1. Gene expression was compared between the HG001 wild type reference strain carrying the pCN51 empty vector (HG001/pCN51) and the HG001 strain carrying a pCN51 derivative with the walRc allele under the control of the Pcad inducible promoter (HG001/pSD3-12). Each condition was done in triplicate and the data were analyzed using the Arraystat software. Genes were considered as differentially expressed if their transcriptional level differed by at least two-fold with a Z-test P-value M-bM-^IM-$ 0.05 in at least two of the replicates.

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High resolution definition of the Staphylococcus aureus WalKR regulon

Project description:High resolution definition of the Staphylococcus aureus WalKR regulon

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Staphylococcus aureus walKR mutants (TPS3130 and TPS3190) vs. parent strains (JKD6009 and JKD6004)

Project description:WalKR is an essential two component regulatory system in S. aureus, thought to control cell wall metabolism. Using genome sequencing of 5 paired clinical isolates of vancomycin-susceptible and vancomycin-intermediate S. aureus we found frequent, but unique, mutations in this locus. To investigate the contribution of these mutations to vancomycin resistance allelic replacement WalK (G223D) and WalR (K208R) mutants were generated and compared to the parent strains. Mutations in walk and walR led to increased vancomycin resistance, reduced biofilms formation and attenuation of virulence, demonstrating that minor genetic changes in this locus can lead to significant changes in bacterial resistance and virulence. Microarray transcriptional comparisons were performed to investigate the regulatory effects of the WalK (G223D) and WalR (K208R) mutations, and demonstrated that while changes in genes affecting cell wall metabolism were detected, more dramatic changes were found in regulation of cellular metabolism. Transcriptional profiling of laboratory derived S. aureus walKR mutants compared to the parent isolates. TPS3130 has a single point mutation in walK, and TPS3190 has a single point mutation in walR. Two condition experiment TPS3130 vs JKD6009 and TPS3190 vs JKD6004. 3 biological replicates per isolate pair, one replicate per slide.

2012-05-31 | E-GEOD-29157 | biostudies-arrayexpress

Staphylococcus aureus walKR mutants (TPS3130 and TPS3190) vs. parent strains (JKD6009 and JKD6004)

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High resolution definition of the Staphylococcus aureus WalKR regulon [RNA-seq]

Project description:High resolution definition of the Staphylococcus aureus WalKR regulon [RNA-seq]

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High resolution definition of the Staphylococcus aureus WalKR regulon [ChIP-seq]

Project description:High resolution definition of the Staphylococcus aureus WalKR regulon [ChIP-seq]

| PRJNA875042 | ENA

High resolution definition of the Staphylococcus aureus WalKR regulon

Project description:This SuperSeries is composed of the SubSeries listed below.

2022-09-01 | GSE212321 | GEO