Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq analysis of MG1655 Escherichia coli (Wild-type and mutant lacking the small regulatory RNA SdsR) experiencing long-term nitrogen starvation in Gutnick minimal media

ABSTRACT: RNA-seq was carried out to compare the transcriptomes of wild-type MG1655 E coli with mutant lacking the prominent sRNA, SdsR, in bacteria that had been exposed to long-term (24hr) nitrogen starvation in Gutnick minimal media. The aim of this was to understand what the regulatory contribution of SdsR was to bacteria experiencing long-term nitrogen starvation.

INSTRUMENT(S): Illumina HiSeq 2000

ORGANISM(S): Escherichia coli K-12

SUBMITTER: Josh McQuail

PROVIDER: E-MTAB-13402 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The airways of cystic fibrosis patients are chronically colonized by a diverse range of microbial pathogens, the composition of which changes throughout life Alteration to the pulmonary environment caused by inter-microbial interactions and pathogen-host interactions influence the type of microbes that can engage in sustained infection. The opportunistic bacterial pathogen Pseudomonas aeruginosa is the primary cause of morbidity and mortality amongst individuals with cystic fibrosis and it is estimated that 60 – 80 % of cystic fibrosis patients experience chronic P. aeruginosa infection by the age of 20 years Aspergillus fumigatus is the most prevalent fungal pathogen isolated from cystic fibrosis airways, affecting up to 58% of patients. It is the causative agent of allergic bronchopulmonary aspergillosis (ABPA), a hypersensitivity disorder resulting from the inhalation of fungal conidia. Although co-colonization of the cystic fibrosis airways by P. aeruginosa and A. fumigatus is rare (3.1 – 15.8 %) disease prognosis is poor when both are present. However, sequential infection is more common. It has recently been suggested that A. fumigatus is more prevalent in juvenile cystic fibrosis patients that has been initially reported due to inconsistencies in the culture methods used to detect A. fumigatus. Despite the prevalence and persistence of A. fumigatus, P. aeruginosa predominates as the primary pathogen in the cystic fibrosis lung, suggesting that interactions with other pathogens such as A. fumigatus may influence the pathogenicity of P. aeruginosa by altering its virulence. We report here an investigation of the effect of culturing P. aeruginosa in the presence of A. fumigatus by measuring differences in growth rate and the overall proteome of the bacteria. It was hypothesized that A. fumigatus creates an environment that promotes a metabolic-driven increase in P. aeruginosa that results in it outcompeting the fungus. The molecular basis of this increased proliferation was investigated further using Label-free quantitative (LFQ) proteomics to characterise the proteome changes in P. aeruginosa when exposed to the supernatant of i) A. fumigatus alone ii) the supernatant of an A. fumigatus/P. aeruginosa co-culture and iii) P. aeruginosa alone. LFQ proteomics involves the simultaneous identification and quantification of thousands of proteins (the ultimate determinants of phenotype) from a single sample has recently been employed to characterise the P. aeruginosa proteome in response to iron limiting conditions, resolving how how the bacteria survives and proliferates in such environments.

Dataset Information

RNA-seq analysis of MG1655 Escherichia coli (Wild-type and mutant lacking the small regulatory RNA SdsR) experiencing long-term nitrogen starvation in Gutnick minimal media

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