Post-transcriptional regulation of gene PA5507 controls Pseudomonas quinolone signal concentration in P. aeruginosa

Mol Microbiol. 2015 May;96(3):670-83. doi: 10.1111/mmi.12963. Epub 2015 Mar 6.

Abstract

Pseudomonas aeruginosa can sense and respond to a myriad of environmental signals and utilizes a system of small molecules to communicate through intercellular signaling. The small molecule 2-heptyl-3-hydroxy-4-quinolone (Pseudomonas Quinolone Signal [PQS]) is one of these signals and its synthesis is important for virulence. Previously, we identified an RpiR-type transcriptional regulator, QapR, that positively affects PQS production by repressing the qapR operon. An in-frame deletion of this regulator caused P. aeruginosa to produce a greatly reduced concentration of PQS. Here, we report that QapR translation is linked to the downstream gene PA5507. We found that introduction of a premature stop codon within qapR eliminates transcriptional autorepression of the qapR operon as expected but has no effect on PQS concentration. This was investigated with a series of lacZ reporter fusions which showed that translation of QapR must terminate at, or close to, the native qapR stop codon in order for translation of PA5507 to occur. Also, it was shown that truncation of the 5' end of the qapR transcript permitted PA5507 translation without translation of QapR. Our findings led us to conclude that PA5507 transcription and translation are both tightly controlled by QapR and this control is important for PQS homeostasis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Artificial Gene Fusion
  • Bacterial Proteins / metabolism*
  • DNA Mutational Analysis
  • Gene Expression Regulation, Bacterial*
  • Genes, Reporter
  • Protein Biosynthesis
  • Pseudomonas aeruginosa / genetics*
  • Pseudomonas aeruginosa / metabolism*
  • Quinolones / metabolism*
  • Transcription Factors / metabolism*
  • beta-Galactosidase / analysis

Substances

  • 2-heptyl-3-hydroxy-4-quinolone
  • Bacterial Proteins
  • Quinolones
  • Transcription Factors
  • beta-Galactosidase