Project description:In this experiment the transcriptional response of the opportunistic human pathogen Pseudomonas aeruginosa to sublethal concentrations of NaClO was investigated. To this aim, four independent cultures of P. aeruginosa PAO1 grown in minimal medium BM2 were treated with NaClO (2 ug/ml) for 1 h at 37 C followed by RNA extraction and microarray analysis. Untreated cultures served as controls.
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. Overall design: The WT (zinc replete) and znuA mutant strain (zinc limited) were grown in cation-defined media to mid-log phase prior to RNA isolation and analysis.
Project description:Oberhardt2008 - Genome-scale metabolic
network of Pseudomonas aeruginosa (iMO1056)
This model is described in the article:
network analysis of the opportunistic pathogen Pseudomonas
Oberhardt MA, Puchałka J, Fryer
KE, Martins dos Santos VA, Papin JA.
J. Bacteriol. 2008 Apr; 190(8):
Pseudomonas aeruginosa is a major life-threatening
opportunistic pathogen that commonly infects immunocompromised
patients. This bacterium owes its success as a pathogen largely
to its metabolic versatility and flexibility. A thorough
understanding of P. aeruginosa's metabolism is thus pivotal for
the design of effective intervention strategies. Here we aim to
provide, through systems analysis, a basis for the
characterization of the genome-scale properties of this
pathogen's versatile metabolic network. To this end, we
reconstructed a genome-scale metabolic network of Pseudomonas
aeruginosa PAO1. This reconstruction accounts for 1,056 genes
(19% of the genome), 1,030 proteins, and 883 reactions. Flux
balance analysis was used to identify key features of P.
aeruginosa metabolism, such as growth yield, under defined
conditions and with defined knowledge gaps within the network.
BIOLOG substrate oxidation data were used in model expansion,
and a genome-scale transposon knockout set was compared against
in silico knockout predictions to validate the model.
Ultimately, this genome-scale model provides a basic modeling
framework with which to explore the metabolism of P. aeruginosa
in the context of its environmental and genetic constraints,
thereby contributing to a more thorough understanding of the
genotype-phenotype relationships in this resourceful and
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Project description:Pseudomonas aeruginosa is an opportunistic human pathogen, infecting immuno-compromised patients and causing persistent respiratory infections in people affected from cystic fibrosis. Pseudomonas strain Pseudomonas aeruginosa PA14 shows higher virulence than Pseudomonas aeruginosa PAO1 in a wide range of hosts including insects, nematodes and plants but the precise cause of this difference is not fully understood. Little is known about the host response upon infection with Pseudomonas and whether or not transcription is being affected as a host defense mechanism or altered in the benefit of the pathogen. In this context the social amoeba Dictyostelium discoideum has been described as a suitable host to study virulence of Pseudomonas and other opportunistic pathogens.
Project description:In this experiment the transcriptional response of the opportunistic human pathogen Pseudomonas aeruginosa towards physiological concentrations of the major human host defense peptide LL-37 was investigated using microarrays. To this aim, three independent cultures of P. aeruginosa PAO1 were grown until mid-log phase in Mueller-Hinton broth and subsequently incubated with either sublethal LL-37 concencentrations (20 µg/ml) or without peptide for 2 h at 37 °C following RNA extraction and microarray analysis.
Project description:The opportunistic pathogen Pseudomonas aeruginosa is among the main colonizers of the lungs of cystic fibrosis (CF) patients. We have isolated and sequenced several P. aeruginosa isolates from the sputum of CF patients and used phenotypic, genomic and proteomic analyses to compare these CF derived strains with each other and with the model strain PAO1.
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:Drosophila harbor substantial genetic variation for antibacterial defense. We allowed wild-caught Drosophila melanogaster to evolve a defense response to systemic infection with the human opportunistic pathogen, Pseudomonas aeruginosa over 10 generations. We performed genome wide transcriptional profiling in selected lines relative to control lines (not infected, but were exposed to the same bottleneck in population size as their paired selected lines by randomly selecting a set of individuals to found the next generation, and then infected at the end of the 10 generations), to identify specifically the genetic basis of the evolved immune response.