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Pseudomonas aeruginosa PA103

ABSTRACT: Pseudomonas aeruginosa T3SS screen

PROVIDER: PRJNA849614 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR19682635.fastq.gz	Fastqsanger.gz
	SRR19682636.fastq.gz	Fastqsanger.gz
	SRR19682637.fastq.gz	Fastqsanger.gz
	SRR19682638.fastq.gz	Fastqsanger.gz
	SRR19682639.fastq.gz	Fastqsanger.gz

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Project description:ClpV3 is a cytoplasmic AAA+ ATPase protein and is an essential component of H3-T6SS in Pseudomonas aeruginosa. Here, we report that an H3-T6SS deletion mutant PAO1(ΔclpV3) significantly affected the virulence-related phenotypes including pyocyanin production, biofilm formation, proteolytic activity and motilities. Most interestingly, the expression of T3SS genes was markedly affected, indicating a strong link between H3-T6SS and T3SS. RNA-Sequencing was performed to globally identify the genes differentially expressed when H3-T6SS was inactivated and the results obtained could be well correlated to the observed phenotypes.

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Project description:The Pseudomonas aeruginosa quorum-sensing (QS) systems contribute to bacterial homeostasis and pathogenicity. Although the AraC family transcription factor VqsM has been characterized to control the production of virulence factors and QS signaling molecules, its detailed regulatory mechanisms still remain elusive. Here, we report that VqsM directly binds to the lasI promoter region, and thus regulates its expression. To identify additional targets of VqsM in P. aeruginosa PAO1, we performed chromatin immunoprecipitation followed by high-throughput DNA sequencing (ChIP-seq) which detected 48 enriched loci harboring VqsM-binding peaks in P. aeruginosa genome. The direct regulation of these genes by VqsM has been confirmed by Electrophoretic mobility shift assays (EMSAs) and quantitative real-time polymerase chain reactions (qRT-PCR). A VqsM-binding motif is found by using MEME suite and verified by footprint assays in vitro. In addition, VqsM directly binds to the promoter regions of antibiotic resistance regulator NfxB and the master type III system regulator ExsA. Notably, the vqsM mutant displayed more resistance to two types of antibiotics and promoted bacterial survival in a mouse model, compared to the wild type PAO1 strain. Collectively, this work provides new cues to better understand the detailed regulatory networks of QS systems, T3SS, and antibiotic resistance. Pseudomonas aeruginosa MAPO1 containing empty pAK1900 or pAK1900-VqsM-VSV

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