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Comparison of the Pseudomonas aeruginosa PA2504 deficient strain and the PAO1161 wild type strain

ABSTRACT: Pseudomonas aeruginosa is an ubiquitous gram-negative bacterium that may colonize a wide range of organisms, including bacteria, plants, and animals. It is a human opportunistic pathogen which shows a great threat to immunocompromised patients. P. aeruginosa displays intrinsic resistance to many antibiotics, and has a high ability to develop novel mechanisms of resistance which forms a threat in hospital environments and makes it extremely hard to eradicate. Additionally over half of the genes of this bacteria have no described function, so it is urgent to search for proteins related to its pathogenicity and antibiotic resistance. The aim of this study was to characterise the P. aeruginosa PA2504 protein of unknown function. Basic phenotypic analysis did not indicate the role of PA2504 in the cell, thus, in order to recognize transcripts affected by the lack of PA2504 transcriptomes of the ΔPA2504 and the wild-type PAO1161 strains were compared using high-throughput RNA sequencing (RNA-seq). Using qRT-PCR method we determined that the level of PA2504 transcript is higher in the stationary phase of growth as compared to the exponential phase of bacterial growth (Log2 FC = 2,77) thus the samples for the RNA-seq experiments were withdrawn from this phase of growth.The RNA-seq revealed that the expression of 42 transcripts was changed in the ΔPA2504 mutant as compared to the parental PAO1161 strain and that the majority of them were connected to the sulphur transport/metabolism.

ORGANISM(S): Pseudomonas aeruginosa

PROVIDER: GSE179150 | GEO | 2021/07/01

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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Comparison of the Pseudomonas aeruginosa PA2504 deficient strain and the PAO1161 wild type strain

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