Burkholderia cenocepacia J2315 exposed to the aminothiol cysteamine during growth in synthetic cystic fibrosis sputum media (SCFM2)
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ABSTRACT: Derived from coenzyme A degradation in mammalian cells, the aminothiol cysteamine may represent a promising new adjunct treatment for pulmonary exacerbations in cystic fibrosis (CF). This experiment aims to characterise the effect of sub-MIC cysteamine on Burkholderia cenocepacia J2315. Samples were cultured in SCFM2 (triplicate), and exposed to either cysteamine or a H2O control.
Project description:Derived from coenzyme A degradation in mammalian cells, the aminothiol cysteamine may represent a promising new adjunct treatment for pulmonary exacerbations in cystic fibrosis (CF). This experiment aims to characterise the effect of sub-MIC cysteamine on P. aeruginosa strain PA14. Samples were cultured in SCFM2 (triplicate), and exposed to either cysteamine or a H2O control.
Project description:Hfq proteins are RNA chaperones that play a critical role in post-transcription regulation of gene expression. Bacteria of the Burkholderia cepacia complex harbor two distinct and functional Hfq proteins, the Hfq and Hfq2. We have previously performed the functional analysis of Hfq and Hfq2 in the pathogen Burkholderia cenocepacia J2315. In order to examine the impacts of each RNA chaperone on the global transcriptome of B. cenocepacia J2315, we performed comparative transcriptome profile of mutants on the hfq and hfq2 genes, using as reference the wild-type strain. For expression profiling, over-night cultures of the Burkholderia cenocepacia J2315 wild-type strain and the isogenic mutants hfq::Tp and M-NM-^Thfq2 grown in LB medium were diluted to an initial OD640 nm of 0.25 into LB medium. Triplicate samples were cultured at 37M-BM-:C with 250 r.p.m. agitation for 16 h and RNA extracted from the three bacterial isolates.
Project description:Burkholderia cenocepacia J2315 exposed to the aminothiol cysteamine during growth in synthetic cystic fibrosis sputum media (SCFM2)
Project description:Hfq proteins are RNA chaperones that play a critical role in post-transcription regulation of gene expression. Bacteria of the Burkholderia cepacia complex harbor two distinct and functional Hfq proteins, the Hfq and Hfq2. We have previously performed the functional analysis of Hfq and Hfq2 in the pathogen Burkholderia cenocepacia J2315. In order to examine the impacts of each RNA chaperone on the global transcriptome of B. cenocepacia J2315, we performed comparative transcriptome profile of mutants on the hfq and hfq2 genes, using as reference the wild-type strain.
Project description:We characterized the transcriptional responses of three S. maltophilia strains during exposure to synthetic CF sputum media (SCFM2) to gain insight into how this organism interacts with the host in the CF lung. These efforts led to the identification of 881 transcripts differentially expressed by all three strains, many of which reflect different metabolic pathways used by S. maltophilia in sputum, and altered stress responses. The latter correlated with increased resistance to peroxide exposure after pre-growth in SCFM2. We also compared the SCFM2 transcriptomes of two S. maltophilia CF isolates with the SCFM2 transcriptome of the acute infection model strain, S. maltophilia K279A, allowing us to identify CF isolate-specific signatures in differential gene expression that may be suggestive of adaptation to the CF lung. Each strain also possessed genes not shared by the other two and here we show that expression of some of the accessory genes in each strain are changed in response to SCFM2. Collectively, this work details the response of S. maltophilia to the CF lung environment, identifying potential survival strategies and metabolic pathways used by S. maltophilia during infections.
Project description:The small RNA ncS35 was deleted from the Burkholderia cenocepacia J2315 genome. Gene expression of mutant was compared to wild type in three growth conditions: exponential phase and stationary phase planktonic growth and biofilm growth, all in LB medium.
Project description:B. cenocepacia J2315 was grown to mid-log phase in different media: LB broth, iso-sensitest broth, basal salt medium with glucose<br>at pH 7 and pH 6, basal salt medium at pH 7 with a reduced iron content, and basal salt medium with glycerol
Project description:Pseudomonas aeruginosa is a major opportunistic human pathogen which employs a myriad of virulence factors. In people with cystic fibrosis (CF) P. aeruginosa frequently colonises the lungs and becomes a chronic infection that evolves to become less virulent over time, but often adapts to favour persistence in the host with alginate-producing mucoid, slow-growing, and antibiotic resistant phenotypes emerging. Cysteamine is an endogenous aminothiol which has been shown to prevent biofilm formation, reduce phenazine production, and potentiate antibiotic activity against P. aeruginosa, and has been investigated in clinical trials as an adjunct therapy for pulmonary exacerbations of CF. Here we demonstrate (for the first time in a prokaryote) that cysteamine prevents glycine utilisation by P. aeruginosa in common with reported activity blocking the glycine cleavage system in human cells. Despite the inhibition of glycine metabolism, cysteamine inhibits hydrogen cyanide (HCN) production by P. aeruginosa, implicating interference in the regulation of virulence. Cysteamine impaired chemotaxis and reductions in pyocyanin and exopolysaccharide production in cysteamine-treated P. aeruginosa and reduced toxicity of secreted factors in a Galleria mellonella model. Thus, cysteamine has additional potent anti-virulence properties targeting P. aeruginosa, further supporting its therapeutic potential in CF and other infections.
Project description:Investigation of gene expression profiles among patients with COPD frequent exacerbations and to find gene targets as predictors of exacerbations