Inhibition of Pseudomonas aeruginosa quorum sensing by chemical induction of the MexEF-oprN efflux pump

Rasmus Kristensen, Jens Bo Andersen, Morten Rybtke, Charlotte Uldahl Jansen, Blaine Gabriel Fritz, Rikke Overgaard Kiilerich, Jesper Uhd, Thomas Bjarnsholt, Katrine Qvortrup, Tim Tolker-Nielsen, Michael Givskov, Tim Holm Jakobsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

The cell-to-cell communication system quorum sensing (QS), used by various pathogenic bacteria to synchronize gene expression and increase host invasion potentials, is studied as a potential target for persistent infection control. To search for novel molecules targeting the QS system in the Gram-negative opportunistic pathogen Pseudomonas aeruginosa, a chemical library consisting of 3,280 small compounds from LifeArc was screened. A series of 10 conjugated phenones that have not previously been reported to target bacteria were identified as inhibitors of QS in P. aeruginosa. Two lead compounds (ethylthio enynone and propylthio enynone) were re-synthesized for verification of activity and further elucidation of the mode of action. The isomeric pure Z-ethylthio enynone was used for RNA sequencing, revealing a strong inhibitor of QS-regulated genes, and the QS-regulated virulence factors rhamnolipid and pyocyanin were significantly decreased by treatment with the compounds. A transposon mutagenesis screen performed in a newly constructed lasB-gfp monitor strain identified the target of Z-ethylthio enynone in P. aeruginosa to be the MexEF-OprN efflux pump, which was further established using defined mex knockout mutants. Our data indicate that the QS inhibitory capabilities of Z-ethylthio enynone were caused by the drainage of intracellular signal molecules as a response to chemical-induced stimulation of the MexEF-oprN efflux pump, thereby inhibiting the autogenerated positive feedback and its enhanced signal-molecule synthesis.
Original languageEnglish
Article numbere0138723
JournalAntimicrobial Agents and Chemotherapy
Volume68
Issue number2
Number of pages25
ISSN0066-4804
DOIs
Publication statusPublished - 2024

Keywords

  • MexEF-oprN
  • Pseudomonas aeruginosa
  • Quorum sensing
  • Quorum-sensing inhibitor
  • Thiol-reactive compound
  • Virulence factors

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