Metabolic profiling of Pseudomonas aeruginosa demonstrates that the anti-sigma factor MucA modulates osmotic stress tolerance.
Metabolic footprinting has shown enormous potential as a phenotyping tool and we are interested in applying it to understand the physiology of the opportunistic pathogen Pseudomonas aeruginosa during its chronic infection of the lungs of cystic fibrosis patients. The selection pressures of surviving in the CF lung environment lead to genetic adaptations of the bacterium. A common adaptation is mutation of the mucA gene, resulting in a loss-of-function mutation to the anti-sigma factor MucA, which leads to a mucoid phenotype as a consequence of the overproduction of the extracellular polysaccharide alginate. However, apart from the mucoid phenotype little is known about the overall metabolic and physiological changes caused by mucA mutation. We investigated the pleiotropic metabolic effects of this mutation using time-resolved metabolic footprinting (extracellular metabolomics), and found changes in the levels of various metabolites associated with osmotic tolerance, including glycine-betaine, trehalose and glutamate. Physiological experiments confirmed that the isogenic mucA22 mutant is less resistant to osmotic stress than the parental PA01 wild-type strain, but only in the stationary phase of growth. Quantitative comparison of the endometabolome of the cells showed differences in the accumulation of osmoprotective metabolites by the wild-type and mucA22 mutant strains, suggesting a switch in osmo-protectant preference from glycine-betaine to trehalose.
Behrends
Volker
V
Biomolecular Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK SW7 2AZ. volker.behrends05@imperial.ac.uk
Ryall
Ben
B
Wang
Xinzhu
X
Bundy
Jacob G
JG
Williams
Huw D
HD
D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media; D003550:Cystic Fibrosis; D016018:Least-Squares Analysis; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular; D009997:Osmotic Pressure / Q000502:physiology; D010641:Phenotype; D025341:Principal Component Analysis; D023302:Protein Footprinting; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012808:Sigma Factor / Q000037:antagonists & inhibitors; D012965:Sodium Chloride / Q000494:pharmacology*; D013312:Stress, Physiological / Q000502:physiology
D001426:Bacterial Proteins; D003470:Culture Media; C107895:MucA protein, Pseudomonas; D012808:Sigma Factor; D012965:Sodium Chloride