Abstract
Pseudomonas aeruginosa coordinates the transcription of hundreds of genes, including many virulence genes, through three hierarchically arranged quorum-sensing (QS) systems, namely las, rhl and pqs. Each system consists of genes involved in autoinducer synthesis, lasI, rhlI and pqsABCDH, as well as cognate-regulatory genes, lasR, rhlR and pqsR. In this study, we analyzed the social behavior of signal-blind (ΔlasR, ΔrhlR, ΔpqsR) and signal-negative (ΔlasI, ΔrhlI, ΔpqsA) mutants from each QS system. As each system controls extracellular common goods but differs in the extent of regulatory control, we hypothesized that all signal-blind mutants can behave as cheaters that vary in their ability to invade a QS-proficient population. We found that lasR and pqsR, but not rhlR, mutants evolve from a wild-type ancestor in vitro under conditions that favor QS. Accordingly, defined lasR and pqsR mutants enriched in wild-type co-culture, whereas rhlR and all signal-negative mutants did not. Both lasR and pqsR mutants enriched with negative frequency dependence, suggesting social interactions with the wild type, although the pqsR mutant also grew well on its own. Taken together, the lasR mutant behaved as a typical cheater, as reported previously. However, the pqsR and rhlR mutants exhibited more complex behaviors, which can be sufficiently explained by positive and negative pleiotropic effects through differential regulation of pqs gene expression in the interconnected QS network. The evolutionary approach adopted here may account for the prevalence of naturally occurring QS mutants.