Project description:The Pae PA1006/nbvF gene was found to be essential for nitrate utilization, biofilm maturation, and virulence. Microarrays were employed to assess global gene expression changes in response to NO3 compared to wild-type. Cells were grown aerobically in the absence or presence of nitrate and the effect on global gene expression was determined; two to three biological replicates for each condition were analyzed.
Project description:In order to understand how Pseudomonas aeruginosa responds to low oxygen we grew strain PAO1 with 3 different oxygen concentrations: 2%, 0.4% and 0% supplemented with nitrate as an electron acceptor. Gene expression under these conditions was compared to that of cells grown with 20% oxygen. Experiment Overall Design: Transcriptome profiles of Pseudomonas aeruginosa cells grown with 20% oxygen were compared to transcriptome profiles obtained by growing cells with 2%, 0.4% and 0% oxygen (with nitrate as the electron acceptor).
Project description:In order to understand how Pseudomonas aeruginosa responds to low oxygen we grew strain PAO1 with 3 different oxygen concentrations: 2%, 0.4% and 0% supplemented with nitrate as an electron acceptor. Gene expression under these conditions was compared to that of cells grown with 20% oxygen. Keywords: Comparison of transcriptome profiles
Project description:Pseudomonas aeruginosa is a virulent opportunistic pathogen responsible for high morbity in COPD, burns , implanted medical devices and cystic fibrosis. Pseudomonas aeruginosa is a problematic colonizer of the human lung. P. aeruginosa produces a phospholipase C (PlcH) that degrades choline-containing lipids such as phosphatidylcholine and sphingomylein that are found in lung surfactant and in host membranes. In this study, we analyzed gene expression in mutants defective in PlcH production (delta-plcH and delta-gbdR) and the wild type when growing in medium with lung surfactant.
Project description:Pseudomonas aeruginosa is a virulent opportunistic pathogen responsible for high morbity in COPD, burns , implanted medical devices and cystic fibrosis. Pseudomonas aeruginosa is a problematic colonizer of the human lung. P. aeruginosa produces a phospholipase C (PlcH) that degrades choline-containing lipids such as phosphatidylcholine and sphingomylein that are found in lung surfactant and in host membranes. In this study, we analyzed gene expression in mutants defective in PlcH production (delta-plcH and delta-gbdR) and the wild type when growing in medium with lung surfactant. Pseudomonas aeruginosa was cultured in liquid cultures with aeration in a defined medium with Survanta, a lung surfactant replacement. Cultures were harvested during mid-exponential phase, and RNA was isolated for microarray analysis. The P. aeruginosa strain PAO1 wild type gene expression was compared to expression profiles from delta-gbdR and delta-plcHR deletion mutants, two mutants defective in PlcH production.
Project description:We report a next-generation sequencing of total RNA from Pseudomonas aeruginosa PAO1 grown in presence of rosmarinic acid (RA) 100mM. Data analysis in comparison with cells grown in absence of RA revealed that the plant compound RA induces a broad transcriptional response in this bacterium, quite similar to the quorum sensing response.
Project description:Opportunistic pathogen Pseudomonas aeruginosa synthesizes a structural homologue of the human alpha2-Macroglobulin protein, a large spectrum protease inhibitor and important player of innate immunity. Delta-MagD and MagD-WT Pseudomonas aeruginosa strains were lysed and lysates submitted to co-immunoprecipitation using anti-MagD antibody (2 biological replicates). Immunoprecipitated proteins were in-gel digested and resulting peptides analysed by nanoLC-MS/MS. Identifications were realised using Mascot and filtered using IRMa software (1% FDR). Results were exported to a relational database (MSIdb) and processed using hEIDI software to identify proteins enriched in MagD-WT samples compared to delta-MagD samples.
Project description:This study addresses the impact of zinc limitation on the opportunistic human pathogen, Pseudomonas aeruginosa. Zinc limitation was assessed in the P. aeruginosa PAO1 strain using an isogenic deletion mutant lacking the periplasmic, zinc solute-binding protein, znuA (PA5498). ZnuA delivers bound zinc to its cognate ABC transporter, ZnuBC, for import into the cytoplasm. Our transcriptional analyses revealed P. aeruginosa to possess a multitude of zinc acquisition mechanisms, each of which were highly up-regulated in the zinc-deficient znuA mutant strain. P. aeruginosa also utilized zinc-independent paralogues of zinc-dependent genes to maintain cellular function under zinc limitation. Together, these data reveal the complex transcriptional response and versatility of P. aeruginosa to zinc depletion.
Project description:In the present study, we employed Affymetrix Pseudomonas aeruginosa GeneChip arrays to investigate the dynamics of global gene expression profiles during the cellular response of Pseudomonas aeruginosa to ortho-phenylphenol, which involved initial growth inhibition and metabolism. Experiment Overall Design: We conducted four independent microarray experiments (biological replicates) in the absence (control) and the presence (experimental) of ortho-phenylphenol. We calculated fold change as the ratio between the signal averages of four untreated (control) and four ortho-phenylphenol-treated (experimental) cultures for 0, 20 and 60 min exposures.
Project description:Microbes in biofilms face the challenge of substrate limitation. In particular, cells in Pseudomonas aeruginosa biofilms growing in the laboratory or during host colonization often become limited for oxygen. Previously we found that phenazines, antibiotics produced by P. aeruginosa, balance the intracellular redox state for cells in biofilms. Here, we show that genes involved in denitrification are induced in phenazine-null (Δphz) mutant biofilms grown under an aerobic atmosphere, even in the absence of nitrate. This finding suggests that resident cells employ a bet-hedging strategy to anticipate the potential availability of nitrate and counterbalance their highly reduced redox state. Consistent with our previous characterization of aerobically-grown colonies supplemented with nitrate, we find that the pathway that is induced in Δphz colonies combines the nitrate reductase activity of the periplasmic enzyme Nap with downstream reduction of nitrite to nitrogen gas catalyzed by the enzymes Nir, Nor, and Nos. This regulatory relationship differs from the denitrification pathway that functions under anaerobic growth with nitrate as the terminal electron acceptor, which depends on the membrane-associated nitrate reductase Nar. We identify sequences in the promoter regions of the nap and nir operons that are required for the effects of phenazines on expression. We also show that specific phenazines have differential effects on nap gene expression. Finally, we provide evidence that individual steps of the denitrification pathway are catalyzed at different depths within aerobically grown biofilms, suggesting metabolic cross-feeding between community subpopulations. (corresponding publication)