Multiple roles of Pseudomonas aeruginosa TBCF10839 PilY1 in motility, transport and infection.

Polymorphonuclear neutrophils are the most important mammalian host defence cells against infections with Pseudomonas aeruginosa. Screening of a signature tagged mutagenesis library of the non-piliated P. aeruginosa strain TBCF10839 uncovered that transposon inactivation of its pilY1 gene rendered the bacterium more resistant against killing by neutrophils than the wild type and any other of the more than 3000 tested mutants. Inactivation of pilY1 led to the loss of twitching motility in twitching-proficient wild-type PA14 and PAO1 strains, predisposed to autolysis and impaired the secretion of quinolones and pyocyanin, but on the other hand promoted growth in stationary phase and bacterial survival in murine airway infection models. The PilY1 population consisted of a major full-length and a minor shorter PilY1* isoform. PilY1* was detectable in small extracellular quinolone-positive aggregates, but not in the pilus. P. aeruginosa PilY1 is not an adhesin on the pilus tip, but assists in pilus biogenesis, twitching motility, secretion of secondary metabolites and in the control of cell density in the bacterial population.

Pseudomonas aeruginosa population biology in chronic obstructive pulmonary disease.

BACKGROUND: The role played by airway infections with Pseudomonas aeruginosa in the course and pathogenesis of chronic obstructive pulmonary disease (COPD) has not yet been resolved. We report on the molecular epidemiology and population biology of P. aeruginosa in COPD. METHODS: P. aeruginosa isolates collected from adults with COPD during a 10-year prospective study were genotyped in 56 binary marker loci of core and accessory genomes. RESULTS: The typing of 134 P. aeruginosa COPD isolates uncovered 60 unrelated bacterial clones. The worldwide dominant clones in the P. aeruginosa population were also the most abundant clones among the COPD isolates. Sporadic or intermittent infection with P. aeruginosa was typical for the airways of patients with COPD. Sequential isolates with the same genotype of the core genome diversified the composition of their accessory genome during the course of the infection. CONCLUSIONS: Intraclonal microevolution and the frequent turnover or loss of clones are typical for infections with P. aeruginosa in COPD. This epidemiological signature differs from that of the chronic carriage of the same P. aeruginosa clone in patients with cystic fibrosis.

Fitness of isogenic colony morphology variants of Pseudomonas aeruginosa in murine airway infection.

Chronic lung infections with Pseudomonas aeruginosa are associated with the diversification of the persisting clone into niche specialists and morphotypes, a phenomenon called 'dissociative behaviour'. To explore the potential of P. aeruginosa to change its morphotype by single step loss-of-function mutagenesis, a signature-tagged mini-Tn5 plasposon library of the cystic fibrosis airway isolate TBCF10839 was screened for colony morphology variants under nine different conditions in vitro. Transposon insertion into 1% of the genome changed colony morphology into eight discernable morphotypes. Half of the 55 targets encode features of primary or secondary metabolism whereby quinolone production was frequently affected. In the other half the transposon had inserted into genes of the functional categories transport, regulation or motility/chemotaxis. To mimic dissociative behaviour of isogenic strains in lungs, pools of 25 colony morphology variants were tested for competitive fitness in an acute murine airway infection model. Six of the 55 mutants either grew better or worse in vivo than in vitro, respectively. Metabolic proficiency of the colony morphology variant was a key determinant for survival in murine airways. The most common morphotype of self-destructive autolysis did unexpectedly not impair fitness. Transposon insertions into homologous genes of strain PAO1 did not reproduce the TBCF10839 mutant morphotypes for 16 of 19 examined loci pointing to an important role of the genetic background on colony morphology. Depending on the chosen P. aeruginosa strain, functional genome scans will explore other areas of the evolutionary landscape. Based on our discordant findings of mutant phenotypes in P. aeruginosa strains PAO1, PA14 and TBCF10839, we conclude that the current focus on few reference strains may miss modes of niche adaptation and dissociative behaviour that are relevant for the microevolution of complex traits in the wild.