Genome-Wide Survey of Pseudomonas aeruginosa PA14 Reveals a Role for the Glyoxylate Pathway and Extracellular Proteases in the Utilization of Mucin

• Published in: Infection and immunity Vol. 85; no. 8
• Main Authors: Flynn, Jeffrey M, Phan, Chi, Hunter, Ryan C
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.08.2017
• Subjects: Acetates - metabolism; Amino Acids - biosynthesis; Bacterial Infections; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biofilms - growth & development; Cystic Fibrosis - microbiology; DNA Transposable Elements - genetics; Genetic Fitness; Genome, Bacterial; Glyoxylates - metabolism; High-Throughput Nucleotide Sequencing; Humans; Metalloendopeptidases - genetics; Metalloendopeptidases - metabolism; Mucins - isolation & purification; Mucins - metabolism; Mutagenesis; Peptide Hydrolases - metabolism; Phenotype; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - growth & development; Pseudomonas aeruginosa - metabolism; Pseudomonas aeruginosa; glyoxylate pathway; cystic fibrosis; mucin; TnSeq
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/iai.00182-17

Chronic airway infections by the opportunistic pathogen are a major cause of mortality in cystic fibrosis (CF) patients. Although this bacterium has been extensively studied for its virulence determinants, biofilm growth, and immune evasion mechanisms, comparatively little is known about the nutrient sources that sustain its growth Respiratory mucins represent a potentially abundant bioavailable nutrient source, although we have recently shown that canonical pathogens inefficiently use these host glycoproteins as a growth substrate. However, given that , particularly in its biofilm mode of growth, is thought to grow slowly , the inefficient use of mucin glycoproteins may be relevant to its persistence within the CF airways. To this end, we used whole-genome fitness analysis, combining transposon mutagenesis with high-throughput sequencing, to identify genetic determinants required for growth using intact purified mucins as a sole carbon source. Our analysis reveals a biphasic growth phenotype, during which the glyoxylate pathway and amino acid biosynthetic machinery are required for mucin utilization. Secondary analyses confirmed the simultaneous liberation and consumption of acetate during mucin degradation and revealed a central role for the extracellular proteases LasB and AprA. Together, these studies describe a molecular basis for mucin-based nutrient acquisition by and reveal a host-pathogen dynamic that may contribute to its persistence within the CF airways.