Global reprogramming of virulence and antibiotic resistance inPseudomonas aeruginosaby a single nucleotide polymorphism in the elongation factor-encoding gene,fusA1

Abstract

AbstractPseudomonas aeruginosais a common opportunistic pathogen. The organism displays elevated intrinsic antibiotic resistance and can cause life-threatening infections. The gene encoding an elongation factor, FusA1, is frequently mutated in clinical isolates ofP. aeruginosafrom patients with cystic fibrosis (CF). Recent work has shown thatfusA1mutants often display elevated aminoglycoside resistance due to increased expression of the aminoglycoside efflux pump, MexXY. In the current work, we isolated a spontaneous gentamicin-resistantfusA1mutant (FusA1P443L) in whichmexXYexpression was increased. Through a combination of proteomic and transcriptomic analyses, we found that thefusA1mutant also exhibited large-scale but discrete changes in the expression of key pathogenicity-associated genes. Most notably, thefusA1mutant displayed greatly increased expression of the Type III Secretion system (T3SS), widely considered to be the most potent virulence factor in theP. aeruginosaarsenal, and also elevated expression of the Type VI Secretion (T6S) machinery. This was unexpected because expression of the T3SS is usually reciprocally coordinated with T6S system expression. ThefusA1mutant also displayed elevated exopolysaccharide production, dysregulated siderophore production, elevated ribosomal protein synthesis, and transcriptomic signatures indicative of translational stress. Each of these phenotypes (and almost all of the transcriptomic and proteomic changes associated with thefusA1mutation) were restored to levels comparable to that in the PAO1-derived progenitor strain by expression of the wild-typefusA1genein trans, indicating that the mutant gene is recessive. Our data show that in addition to elevating antibiotic resistance throughmexXYexpression (although we also identify additional contributory resistance mechanisms), mutations infusA1can lead to highly-selective dysregulation of virulence gene expression.