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The small RNAs PA2952.1 and PrrH as regulators of virulence, motility and iron metabolism in Pseudomonas aeruginosa

ABSTRACT: Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that undergoes swarming motility in response to semisolid conditions with amino acids as a nitrogen source. With a genome encoding hundreds of potential intergenic small RNAs (sRNAs), P. aeruginosa can easily adapt to different conditions and stresses. Overexpression of the previously uncharacterized PA2952.1 resulted in decreased swarming and swimming motility, increased gentamicin and tobramycin resistance under swarming conditions, and increased trimethoprim susceptibility. RNA-Seq and proteomics were performed on the wildtype overexpressing PA2952.1 cf. the empty vector control under swarming conditions using TMT10 labeled differential protein expression analysis. The TMT10 reporter channels were sequentially assigned in increasing reporter mass as TMT0..TMT9. We assigned four TMT10 channels (TMT0..TMT3) to biological replicate samples from the EV strain, and three TMT10 channels (TMT4..TMT6) to biological replicate samples from the PA2952.1 strain. Data was acquired over a single 180-minute HPLC-MS run, with peptides identified using X!tandem (cyclone 2012.10.01.1). Protein level expression values were computed using the sum of the reporter channel values from their member peptides, with protein assignment handled by X!tandem. Expression values were transformed into a log2 scale for differential analysis.

INSTRUMENT(S): Orbitrap Q Exactive HF-X

ORGANISM(S): Pseudomonas Aeruginosa

SUBMITTER: Robert E.W. Hancock

PROVIDER: MSV000085955 | MassIVE | Sat Aug 15 18:25:00 BST 2020

REPOSITORIES: MassIVE

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