Identification of a novel RhlI/R-PrrH-LasI/Phzc/PhzD signalling cascade and its implication in P. aeruginosa virulence

Emerg Microbes Infect. 2019;8(1):1658-1667. doi: 10.1080/22221751.2019.1687262.

Abstract

Small regulatory RNAs (sRNAs) act as key regulators in many bacterial signalling cascades. However, in P. aeruginosa, the sRNAs involved in quorum sensing (QS) regulation and their function are still largely unknown. Here, we explored how the prrH locus sRNA influences P. aeruginosa virulence in the context of the QS regulatory network. First, gain- and loss-of-function studies showed that PrrH affects pyocyanin, elastase and rhamnolipid production; biofilm formation; and swimming and swarming motility and impaired the viability of P. aeruginosa in human whole blood. Next, our investigation disclosed that LasI and PhzC/D were directly repressed by PrrH. In addition, RhlI, the key member of the rhl QS system, diminished the expression of PrrH and enhanced the expression of downstream genes. Bioinformatics analysis found two binding sites of RhlR, the transcription factor of the rhl system, on the promoter region of prrH. Further β-galactosidase reporter and qPCR assays confirmed that PrrH was transcriptionally repressed by RhlR. Collectively, our data identified a novel RhlI/R-PrrH-LasI/PhzC/PhzD regulatory circuitry that may contribute to P. aeruginosa pathogenesis. Our findings indicate that PrrH is a quorum regulatory RNA (Qrr) in P. aeruginosa and provide new insight into PrrH's function.

Keywords: P. aeruginosa; PrrH; quorum sensing; sRNA; virulence.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Gene Expression Regulation, Bacterial*
  • Humans
  • Pseudomonas Infections / microbiology*
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / metabolism
  • Pseudomonas aeruginosa / pathogenicity*
  • Quorum Sensing
  • RNA, Bacterial / genetics
  • RNA, Bacterial / metabolism*
  • Virulence

Substances

  • Bacterial Proteins
  • LasI protein, Pseudomonas aeruginosa
  • RNA, Bacterial

Grants and funding

This work was supported by the National Natural Science Foundation of China (grant number 81702063, 81871703); the Guangzhou Science Technology and Innovation Commission (grant number 201707010296); the Natural Science Foundation of Guangdong Province (grant number 2015A030310439); Guangdong Provincial Hospital of Traditional Chinese Medicine (grant number YN2018QJ01).