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Transcriptional analysis of transposon mutants disrupted in glutathione biosynthesis in Acinetobacter baumannii

ABSTRACT: Transcriptional analysis of transposon mutants disrupted in glutathione biosynthesis in Acinetobacter baumannii

PROVIDER: PRJNA793888 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17414737_1.fastq.gz	Fastqsanger.gz
	SRR17414737_2.fastq.gz	Fastqsanger.gz
	SRR17414738_1.fastq.gz	Fastqsanger.gz
	SRR17414738_2.fastq.gz	Fastqsanger.gz
	SRR17414739_1.fastq.gz	Fastqsanger.gz

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Transcriptional analysis of transposon mutants disrupted in glutathione biosynthesis in Acinetobacter baumannii

Project description:In this study we compare logrithmically grown Acinetobacter baumannii wildtype to a transposon mutant that is disrupted in gshA, the first step in glutathione biosynthesis using RNASeq to identify novel pathways where glutathione may be involved.

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Transcriptional analysis of transposon mutants disrupted in glutathione biosynthesis in Pseudomonas aeruginosa

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Transcriptional analysis of transposon mutants disrupted in glutathione biosynthesis in Pseudomonas aeruginosa

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Transcriptional analysis of transposon mutants disrupted in bacillithiol biosynthesis in Staphylococcus aureus

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Project description:In this study we compare logrithmically grown Staphylococcus aureus SAUSA300 wildtype to a transposon mutant that is disrupted in bshC, the third step in bacillithiol biosynthesis using RNASeq to identify novel pathways where bacillithiol may be involved.

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Project description:A library of transposon mutants were generated in S.aureus strain SH1000 using a custom Mariner transposon with an outward-facing T7 promoter. Genomic DNA was extracted from the library and digested with an appropriate restriction enzyme (AluI or RsaI). Labelled RNA run-offs were produced from the T7 promoter and hybridised to a tiling microarray to determine the position of the mutant. This was done using a library of a million S. aureus mutants, and genes not disrupted by a transposon were inferred to be essential for replication and growth in vitro.

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Pyochelin methylation by S. aureus transposon library mutants

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Project description:Pseudomonas aeruginosa infections for individuals with Cystic Fibrosis (CF), result in high morbidity and mortality, with premature death often occurring. These infections are complicated by the formation of biofilms in the sputum. Antibiotic therapy is stymied by antibiotic resistance of the biofilm matrix, making novel anti-biofilm strategies highly desirable. Within the P. aeruginosa biofilm, the redox factor pyocyanin enhances biofilm integrity by intercalating with extracellular DNA. The antioxidant glutathione (GSH) reacts with pyocyanin to disrupt intercalation. This study investigated GSH disruption by assaying the physiological effects of GSH and DNase I on biofilms of clinical CF isolates grown in artificial CF sputum media (ASMDM+). Confocal scanning laser microscopy showed that 2mM GSH alone or combined with DNase I significantly disrupted the immature (24 hour) biofilms of Australian Epidemic Strain (AES) isogens AES-1R and AES-1M. GSH alone greatly disrupted the mature (72 hour) biofilm of AES-1R, resulting in significant differential expression of 587 genes, as evidenced by RNA-sequencing. Upregulated systems included cyclic diguanylate and pyoverdine biosynthesis, the Type VI secretion system, nitrate metabolism and translational machinery. Physiochemical biofilm disruption with GSH revealed a metabolically active cellular physiology distinct from either mature or dispersed biofilm physiology. RNA-seq results were validated by biochemical assay and qPCR. Biofilms of a range of CF isolates disrupted with GSH and DNase I were significantly more susceptible to ciprofloxacin, and increased antibiotic effectiveness was achieved at 10mM GSH. This study demonstrated that GSH alone or with DNase I represent effective anti-biofilm treatments when combined with appropriate antibiotics.

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