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ABSTRACT: Tn-seq of Salmonella Typhimurium under iron-restriction with oxidative stressor

PROVIDER: PRJNA705518 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13809427.fastq.gz	Fastqsanger.gz

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Project description:Tn-seq of Salmonella Typhimurium under iron-restriction with oxidative stressor

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Project description:Salmonella enterica serovar Typhimurium (S. Typhimurium) is a zoonotic pathogen that causes diarrheal disease in humans and animals. During salmonellosis, S. Typhimurium colonizes epithelial cells lining the gastrointestinal tract. S. Typhimurium has an unusual lifestyle in epithelial cells that begins within an endocytic-derived Salmonella-containing vacuole (SCV), followed by escape into the cytosol, epithelial cell lysis and bacterial release. The cytosol is a more permissive environment than the SCV and supports rapid bacterial growth. The physicochemical conditions encountered by S. Typhimurium within the cytosol, and the bacterial genes required for cytosolic colonization, remain unknown. Here we have exploited the parallel colonization strategies of S. Typhimurium in epithelial cells to decipher the two niche-specific bacterial virulence programs. By combining a population-based RNA-seq approach with single-cell microscopic analysis, we identified bacterial genes/sRNAs with cytosol-specific or vacuole-specific expression signatures. Using these genes/sRNAs as environmental biosensors, we defined that Salmonella is exposed to iron and manganese deprivation and oxidative stress in the cytosol and zinc and magnesium deprivation in the SCV. Furthermore, iron availability was critical for optimal S. Typhimurium replication in the cytosol, as well as entC, fepB, soxS and sitA-mntH. Virulence genes that are typically associated with extracellular bacteria, namely Salmonella pathogenicity island 1 (SPI1) and SPI4, had a cytosolic-specific expression profile. Our study reveals that the cytosolic and vacuolar S. Typhimurium virulence gene programs are unique to, and tailored for, residence within distinct intracellular compartments. Therefore, this archetypical vacuole-adapted pathogen requires extensive transcriptional reprogramming to successfully colonize the mammalian cytosol.

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Salmonella enterica subsp. enterica serovar Typhimurium str. 14028S

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Project description:HilD is a regulator of Salmonella pathogenicity island 1 (SPI-1) virulence genes in Salmonella enterica serovar Typhimurium. To identify novel HilD-regulated genes, we mapped the genome-wide association of HilD in S. Typhimurium under SPI-1-inducing conditions (high salt, low aeration) using ChIP-seq. HilD was C-terminally tagged with 3 FLAG tags in strain 14028s.

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Project description:We report the findings of genes under the transcriptional control of Gre factors by comparing RNA-seq data from parental, wild-type Salmonella enterica enterica Typhimurium 14028s and a ∆greA ∆greB strain. By comparing differential gene expression, we identify a metabolic scheme for responding to oxidative stress that is regulated in part by the action of Gre factors

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Project description:PacBio SMRT-seq of wild-type, ∆metJ, and ∆yhdJ Salmonella enterica serovar Typhimurium grown under SPI-1-inducing condition.

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