Abstract
Bacteriophages play an important role in bacterial virulence and phenotypic variation. It has been shown that filamentous bacteriophage Pf4 of Pseudomonas aeruginosa strain PAO1 mediates the formation of small-colony variants (SCVs) in biofilms. This morphology type is associated with parameters of poor lung function in cystic fibrosis patients, and SCVs are often more resistant to antibiotics than wild-type cells. P. aeruginosa strain PA14 also contains a Pf1-like filamentous prophage, which is designated Pf5, and is highly homologous to Pf4. Since P. aeruginosa PA14 produces SCVs very efficiently in biofilms grown in static cultures, the role of Pf5 in SCV formation under these conditions was investigated. The presence of the Pf5 replicative form in total DNA from SCVs and wild-type cells was detected, but it was not possible to detect the Pf5 major coat protein by immunoblot analysis in PA14 SCV cultures. This suggests that the Pf5 filamentous phage is not present at high densities in the PA14 SCVs. Consistent with these results, we were unable to detect coaB expression in SCV cultures and SCV colonies. The SCV variants formed under static conditions were not linked to Pf5 phage activity, since Pf5 insertion mutants with decreased or no production of the Pf5 RF produced SCVs as efficiently as the wild-type strain. Finally, analysis of 48 clinical P. aeruginosa isolates showed no association between the presence of Pf1-like filamentous phages and the ability to form SCVs under static conditions; this suggests that filamentous phages are generally not involved in the emergence of P. aeruginosa SCVs.
Highlights
Pseudomonas aeruginosa is a versatile bacterium that can inhabit many environments, including soil, marshes, plants and water
Recent work has indicated that phenotypic variants, such as P. aeruginosa small-colony variant (SCV), play an important role in biofilm formation
We focused on the relationship between SCVs produced by P. aeruginosa strain PA14, and the presence of a Pf1-like filamentous bacteriophage in the PA14 genome
Summary
Pseudomonas aeruginosa is a versatile bacterium that can inhabit many environments, including soil, marshes, plants and water. P. aeruginosa predominantly grows in organized communities called biofilms. P. aeruginosa grows as a biofilm in the lungs of cystic fibrosis (CF) patients, and, despite long-term antibiotic treatment, the lungs remain infected (Hoiby et al, 2001; Singh et al, 2000). Recent work has indicated that phenotypic variants (bacteria that show a stable change in phenotype whether genetic or epigenetic), such as P. aeruginosa SCVs, play an important role in biofilm formation Hyperadherent antibiotic-resistant SCVs appear after treatment of P. aeruginosa PA14 cultures with the antibiotic kanamycin (Drenkard & Ausubel, 2002)
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