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Determination of the RNA interactome of the RNA-binding protein Hfq across three Pseudomonas aeruginosa strains

ABSTRACT: Determination of the RNA interactome of the RNA-binding protein Hfq across three Pseudomonas aeruginosa strains

PROVIDER: PRJNA718256 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14094300.fastq.gz	Fastqsanger.gz
	SRR14094301.fastq.gz	Fastqsanger.gz
	SRR14094302.fastq.gz	Fastqsanger.gz
	SRR14094303.fastq.gz	Fastqsanger.gz
	SRR14094304.fastq.gz	Fastqsanger.gz

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Determination of the RNA interactome of the RNA-binding protein Hfq across three Pseudomonas aeruginosa strains

Project description:Determination of the RNA interactome of the RNA-binding protein Hfq at three different time-point during growth on the three strains PAO1, PA14 and IHMA87, representing the three major P. aeruginosa phylogenetic lineages, using RIP-seq.

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Determination of sRNAs interacting with vfr mRNA across three Pseudomonas aeruginosa strains

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The global RNA-RNA interactome of Klebsiella pneumoniae unveils a small RNA regulator of cell division (dRNA-Seq)

Project description:The ubiquitous RNA chaperone Hfq is involved in the regulation of key biological processes in many species across the bacterial kingdom. In the opportunistic human pathogen Klebsiella pneumoniae, deletion of the hfq gene affects the global transcriptome, virulence, and stress-resistance, however, the ligands of the major RNA-binding protein in this species have remained elusive. In this study, we have combined transcriptomic, co-immunoprecipitation and global RNA interactome analyses to compile an inventory of conserved and species-specific RNAs bound by Hfq, and to monitor Hfq-mediated RNA-RNA interactions. In addition to dozens of previously unknown RNA-RNA pairs, our study revealed an Hfq-dependent sRNA, DinR, which is processed from the 3’ terminal portion of dinI mRNA. Transcription of dinI is controlled by the master regulator of the SOS response, LexA. As DinR accumulates in K. pneumoniae in response to DNA damage, the sRNA represses translation of the ftsZ transcript by occupation of the ribosome binding site. Ectopic overexpression of DinR causes depletion of ftsZ mRNA and inhibition of cell division, while deletion of dinR antagonizes cell elongation in the presence of DNA damage. Collectively, our work highlights the important role of RNA-based gene regulation in K. pneumoniae and uncovers the central role of DinR in LexA-controlled cell cycle inhibition during the SOS response.

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The global RNA-RNA interactome of Klebsiella pneumoniae unveils a small RNA regulator of cell division (RIP-Seq)

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PAO1 Hfq-V RNA-seq

Project description:A key challenge for understanding the role(s) played by short, non-coding RNAs (sRNAs) in bacteria is identifying the mRNA targets regulated by the sRNAs. Because the Hfq protein mediates the interactions between many sRNAs and the corresponding target mRNAs, one apporach to identify the mRNA targets of sRNAs is to capture sRNA:mRNA interactions occuring on Hfq by exposing cells to UV-irradiation, which forms cross-links between nucleic acids and proteins. We subjected cells of P. aeruginosa strain PAO1 and a derivative of PAO1 harboring a C-terminal VSV-G epitope on Hfq to UV-irradiation, immune-precipitated the Hfq-RNA complexes, ligated neighboring RNA molecules together with RNA Ligase, and then purififed the resulting RNAs. These RNAs were converted into cDNA libraries and sequenced using the Illumina NextSeq platform and then subjected to RIL-seq analysis pipeline (version 0.78) to identify chimeric RNA molecules. We also performed RNA-seq for PAO1 ∆phrS cells harboring an empty vector (pEV) or a vector expressing PhrS (pPhrS).

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