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The Lrp/AsnC-type regulator PA2577 controls the Eam-like transporter gene PA2576 in Pseudomonas aeruginosa (ChIP-seq)

ABSTRACT: Pseudomonas aeruginosa is a facultative human pathogen especially dangerous for immunocompromised patients. It is known from its high adaptability and complex regulatory systems involving huge repertoire of transcriptional regulators (TRs). The Lrp/AsnC family consists of proteins engaged mostly in regulation of processes such as amino acid metabolism but the function of most of them remains unknown. The aim of this study was characterization of the Lrp/AsnC-type regulator PA2577 of P. aeruginosa. Transcriptional profiling of P. aeruginosa PAO1161 overexpressing PA2577 revealed changes in mRNA level of 171 genes. Among downregulated loci PA2576 gene, encoding an EamA-like membrane transporter transcribed divergently to PA2577, was identified. ChIP-seq analysis together with EMSA analysis revealed binding site of PA2577 in PA2577/PA2576 intergenic region, indicating direct regulation of PA2576 expression by PA2577. PA2576 was shown to interact with PA5006 and PA3694 proteins predicted to be involved in membrane processes, especially LPS biosynthesis. Defects of growth of PA2577 and PA2576 deficient cells in conditions of polymyxin B supplementation was shown. Overall, presented data uncovered the regulatory properties of PA2577 acting as the repressor of divergently transcribed PA2576 gene, being a part of LPS biogenesis process in P. aeruginosa.

ORGANISM(S): Pseudomonas aeruginosa

PROVIDER: GSE186746 | GEO | 2021/12/10

REPOSITORIES: GEO

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Project description:Representatives of two families of bacterial Par proteins, ParA and ParB, are encoded by the majority of bacterial chromosomes in the close vicinity of oriC. ParA(Soj) and ParB(Spo0J) proteins of Pseudomonas aeruginosa are both important for optimal growth, nucleoids segregation, cell division and different types of motility. Comparative transcriptome analysis of parAnull, parBnull mutants versus parental PAO1161 strain of P. aeruginosa demonstrated global changes in genes expression pattern in logarithmic phase of planktonic cultures grown on rich medium. The set of genes that were similarly regulated in both mutant strains as compared to the wild-type strain as well as two sets of genes uniquely affected in the particular mutant were defined suggesting that ParA and ParB may act in common and independently. In general, many genes involved in cell division, DNA and RNA processing and metabolic processes were down-regulated in mutant cells, in contrast genes which products play a role in adaptation, protection, motility, cell-to-cell signaling as well as signal transduction increased their expression in par mutant cells. Besides their role in chromosome segregation, ParA and ParB seem to have the potential to regulate genes transcription. The altered expression of a large number of genes encoding known or predicted transcriptional regulators and genes coding for products involved in c-di-GMP signalling, suggests that the part of observed global changes in genes expression pattern in parAnull and parBnull mutants might be the effect of indirect regulation mediated by regulatory genes under ParA and ParB control. The extended regulatory network provides the mechanism to modulate genes expression in response to the stage of the chromosome segregation process and cell cycle. Pseudomonas aeruginosa PAO1161 (leu, r-, RifR), derivative of PAO1, as a control (reference) strain, Pseudomonas aeruginosa PAO1161 parA1-40::smh (parAnull) and Pseudomonas aeruginosa PAO1161 parB1-18::TcR (parBnull) disruption mutant strains were used in the experiments. Three independent biological replicates of total RNA were isolated for each strain from logarithmic (Log) phase of planktonic culture grown on rich medium (L broth) at 37oC. In total, nine samples of RNA were prepared.

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The Lrp/AsnC-type regulator PA2577 controls the Eam-like transporter gene PA2576 in Pseudomonas aeruginosa (ChIP-seq)

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