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PA3027, a novel AraC-type transcriptional regulator activating genes of glycerolipid metabolism in Pseudomonas aeruginosa (PA3027 overexpression RNA-seq)

ABSTRACT: Pseudomonas aeruginosa encodes a large set of transcriptional regulators which allow to modulate and manage cellular metabolism to survive in variable environmental conditions, including the human body. The AraC family regulators are an abundant group of transcriptional regulators in bacteria, mostly acting as gene expression activators controlling diverse cellular functions e.g. carbon metabolism, stress response and virulence. PA3027 protein from P. aeruginosa has been classified in silico as a putative AraC-type transcriptional regulator. Comparative transcriptome analysis of P. aeruginosa PAO1161 overexpressing PA3027 versus empty vector control revealed a spectacular over 15-fold up-regulation of expression of pa3026-pa3024, PA3464 and PA3342 genes potentially involved in glycerolipid metabolism. Concomitantly, the ChIP-seq analysis using FLAG-PA3027 under mild overproduction conditions revealed at least 22 PA3027 binding sites in the PAO1161 genome, including promoter regions of genes showing a major increase in expression in response to PA3027 excess. The presented data showed that PA3027 acts as the transcriptional regulator in P. aeruginosa activating genes engaged in glycerolipid metabolism.

ORGANISM(S): Pseudomonas aeruginosa

PROVIDER: GSE163555 | GEO | 2021/05/11

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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PA3027, a novel AraC-type transcriptional regulator activating genes of glycerolipid metabolism in Pseudomonas aeruginosa (PA3027 overexpression RNA-seq)

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