Characterization of the Entner-Doudoroff pathway in Pseudomonas aeruginosa catheter-associated urinary tract infections

• Published in: Journal of bacteriology 2024-01, Vol.206 (1), p.e0036123-e0036123
• Main Authors: El Husseini, Nour, Mekonnen, Solomon A, Hall, Cherisse L, Cole, Stephanie J, Carter, Jared A, Belew, Ashton T, El-Sayed, Najib M, Lee, Vincent T
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Bacteriology; Carbon; Carbon sources; Catheters; Citric acid; Complex media; Defects; Disease Models, Animal; Entner-Doudoroff pathway; Gene regulation; Gene sequencing; Genes; Gluconates; Glucose; Glucose - metabolism; Glyceraldehyde; Laboratory animals; Medical instruments; Mice; Mutants; Mutation; Nosocomial infection; Opportunist infection; Pathogens; Phenotypes; Pseudomonas aeruginosa; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - metabolism; Pseudomonas Infections; Pyruvic acid; Quorum sensing; Research Article; Tricarboxylic acid cycle; Urea; Urinary tract; Urinary tract diseases; Urinary Tract Infections; Urine; Urogenital system; Virulence
• ISSN: 0021-9193; 1098-5530
• DOI: 10.1128/jb.00361-23

is an opportunistic nosocomial pathogen responsible for a subset of catheter-associated urinary tract infections (CAUTI). In a murine model of CAUTI, we previously demonstrated that urea within urine suppresses quorum sensing and induces the Entner-Doudoroff (E-D) pathway. The E-D pathway consists of the genes , , , and . Zwf and Pgl convert glucose-6-phosphate into 6-phosphogluconate. Edd hydrolyzes 6-phosphogluconate to 2-keto-3-deoxy-6-phosphogluconate (KDPG). Finally, Eda cleaves KDPG to glyceraldehyde-3-phosphate and pyruvate, which enters the citric acid cycle. Here, we generated in-frame E-D mutants in the strain PA14 and assessed their growth phenotypes on chemically defined and complex media. These E-D mutants have a growth defect when grown on glucose or gluconate as the sole carbon source, which is similar to results previously reported for PAO1 mutants lacking E-D genes. RNA-sequencing following short exposure to urine revealed minimal gene regulation differences compared to the wild type. In a murine CAUTI model, virulence testing of E-D mutants revealed that two mutants lacking and showed minor fitness defects. Infection with the ∆ strain exhibited a 20% increase in host survival, and the ∆ strain displayed decreased colonization of the catheter and kidneys. Consequently, our findings suggest that the E-D pathway in is dispensable in this model of CAUTI. IMPORTANCE Prior studies have shown that the Entner-Doudoroff pathway is up-regulated when is grown in urine. Pseudomonads use the Entner-Doudoroff (E-D) pathway to metabolize glucose instead of glycolysis, which led us to ask whether this pathway is required for urinary tract infection. Here, single-deletion mutants of each gene in the pathway were tested for growth on chemically defined media with single-carbon sources as well as complex media. The effect of each mutant on global gene expression in laboratory media and urine was characterized. The virulence of these mutants in a murine model of catheter-associated urinary tract infection revealed that these mutants had similar levels of colonization indicating that glucose is not the primary carbon source utilized in the urinary tract.