GlpR Regulates Motility and Viscoelasticity Properties of Pseudomonas aeruginosa

Despite the onset of highly effective modulator therapies people with Cystic Fibrosis continue to experience recurrent microbial lung infections. Many of these individuals will have at least one positive culture per year for Pseudomonas aeruginosa, a bacterium that readily adapts to live in the lungs. Of these adaptations, the formation of a protective biofilm and changes in motility are hallmarks of established infections. We have shown previous evidence that glycerol metabolism and the P. aeruginosa glycerol regulon repressor, GlpR, is linked to enhanced biofilm production and reduced susceptibility to tobramycin. We report that loss of GlpR contributes to higher viscosity and elasticity in synthetic cystic fibrosis sputum media. Further, we show that the loss of the glycerol repressor GlpR, or growth on glycerol, both resulting in derepression of the GlpR/glycerol regulon, cause decreased motility in both acute and chronic CF-adapted lab strains. RNA sequencing analysis indicated that loss of GlpR altered the expression of genes involved in motility, iron scavenging, transport, metabolism, and virulence. An in silico search of P. aeruginosas genome using GlpRs previously determined binding consensus site identified potential bindings sites in genes related to biofilm development, motility, antibiotic resistance, and metabolism, and these binding sites were confirmed using chromatin immunoprecipitation sequencing. Collectively, our results indicate that GlpR regulates P. aeruginosa phenotypes that facilitate persistence in the CF airway and we provide evidence that GlpR regulates genes outside of the canonical glp regulon.