The Pseudomonas aeruginosa RhlR-controlled aegerolysin RahU is a low-affinity rhamnolipid-binding protein

Abstract

The opportunistic pathogen Pseudomonas aeruginosa uses quorum-sensing systems to regulate collective behaviour in response to the environment, by linking the expression of particular genes to population density. The quorum-sensing transcription factors LasR and RhlR and their cognate N-acyl-homoserine lactone (HSL) signals N-3-oxo-dodecanoyl-L-HSL (3OC12-HSL) and N-butanoyl-L-HSL (C4-HSL) control the expression of several hundred genes, which include those involved in virulence and biofilm formation. Here, we have focused on regulation of the expression of the putative virulence factor gene, rahU. We show that the intact las-rhl box immediately upstream of the -35 promoter element is needed for rahU expression in P. aeruginosa. Using β-galactosidase assays and quantification of the mRNA levels for rahU, lasR and rhlR, we provide evidence that for rahU promoter activity, 3OC12-HSL-LasR is not sufficient, and instead C4-HSL-RhlR is the trigger. Furthermore, surface plasmon resonance analysis revealed that RahU binds the biosurfactant rhamnolipids. Thus, this is the first report of a bacterial molecule that interacts with RahU.