Complete Genome Sequence of Pseudomonas aeruginosa Reference Strain PAK

Cain, Amy K.; Nolan, Laura M.; Sullivan, Geraldine J.; Whitchurch, Cynthia B.; Filloux, Alain; Parkhill, Julian

doi:10.1128/mra.00865-19

Cited by 29 publications

(16 citation statements)

References 16 publications

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“…Homologues of proteins involved in natural transformation were identified in P. aeruginosa PAO1 using blast p [ 31 ]. The Pseudomonas.com resource [ 32 ] and the PAK genome [ 33 ] were used to identify P. aeruginosa orthologues.…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas aeruginosa is capable of natural transformation in biofilms

et al. 2020

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Natural transformation is a mechanism that enables competent bacteria to acquire naked, exogenous DNA from the environment. It is a key process that facilitates the dissemination of antibiotic resistance and virulence determinants throughout bacterial populations. Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen that produces large quantities of extracellular DNA (eDNA) that is required for biofilm formation. P. aeruginosa has a remarkable level of genome plasticity and diversity that suggests a high degree of horizontal gene transfer and recombination but is thought to be incapable of natural transformation. Here we show that P. aeruginosa possesses homologues of all proteins known to be involved in natural transformation in other bacterial species. We found that P. aeruginosa in biofilms is competent for natural transformation of both genomic and plasmid DNA. Furthermore, we demonstrate that type-IV pili (T4P) facilitate but are not absolutely essential for natural transformation in P. aeruginosa .

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Section: Methodsmentioning

confidence: 99%

Pseudomonas aeruginosa is capable of natural transformation in biofilms

et al. 2020

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“…Homologs of proteins involved in natural transformation were identified in P. aeruginosa PAO1 using BLASTp 28 . The Pseudomonas.com resource 29 and the PAK genome 30 were used to identify P. aeruginosa orthologs.…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas aeruginosa is capable of natural transformation in biofilms

Nolan

Turnbull

Katrib

et al. 2019

Preprint

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Natural transformation is a mechanism that enables competent bacteria to acquire naked, exogenous DNA from the environment. It is a key process that facilitates the dissemination of antibiotic resistance and virulence determinants throughout bacterial populations. Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen that produces large quantities of extracellular DNA (eDNA) that is required for biofilm formation. P. aeruginosa has a remarkable level of genome plasticity and diversity that suggests a high degree of horizontal gene transfer and recombination but is thought to be incapable of natural transformation. Here we show that P. aeruginosa possesses homologs of all proteins known to be involved in natural transformation in other bacterial species. We found that P. aeruginosa in biofilms is competent for natural transformation of both genomic and plasmid DNA. Furthermore, we demonstrate that type IV pili (T4P) facilitate but are not absolutely essential for natural transformation in P. aeruginosa.

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“…To investigate the effects of AlyP1400 on a non-mucoid strain of P . aeruginosa , the flow cell biofilm of lab strain PAK [ 18 ] was cultivated and treated as described above for CF27. CFU counting on LB agar plates of the dispersal cells from various treatments were carried out using serially diluted PAK dispersal cells.…”

Section: Methodsmentioning

confidence: 99%

Characterizations of the viability and gene expression of dispersal cells from Pseudomonas aeruginosa biofilms released by alginate lyase and tobramycin

2021

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Biofilm infections are hard to manage using conventional antibiotic treatment regimens because biofilm structures discourage antibiotics from reaching the entire bacterial community and allow pathogen cells to persistently colonize and develop a plethora of tolerance mechanisms towards antibiotics. Moreover, the dispersed cells from biofilms can cause further complications by colonizing different sites and establishing new cycles of biofilms. Previously, we showed that alginate lyase enzyme (AlyP1400), purified from a marine Pseudoalteromonas bacterium, reduced Pseudomonas aeruginosa biofilm biomass and boosted bactericidal activity of tobramycin by degrading alginate within the biofilm extracellular polymeric substances matrix. In this work, we used a flow cytometry-based assay to analyze collected dispersal cells and demonstrated the synergy between tobramycin with AlyP1400 in enhancing the release of both live and dead biofilm cells from a mucoid P. aeruginosa strain CF27, which is a clinical isolate from cystic fibrosis (CF) patients. Interestingly, this enhanced dispersal was only observed when AlyP1400 was combined with tobramycin and administered simultaneously but not when AlyP1400 was added in advance of tobramycin in a sequential manner. Moreover, neither the combined nor sequential treatment altered the dispersal of the biofilms from a non-mucoid P. aeruginosa laboratory strain PAK. We then carried out the gene expression and tobramycin survival analyses to further characterize the impacts of the combined treatment on the CF27 dispersal cells. Gene expression analysis indicated that CF27 dispersal cells had increased expression in virulence- and antibiotic resistance-related genes, including algR, bdlA, lasB, mexF, mexY, and ndvB. In the CF27 dispersal cell population, the combinational treatment of AlyP1400 with tobramycin further induced bdlA, mexF, mexY, and ndvB genes more than non-treated and tobramycin-treated dispersal cells, suggesting an exacerbated bacterial stress response to the combinational treatment. Simultaneous to the gene expression analysis, the survival ability of the same batch of biofilm dispersal cells to a subsequent tobramycin challenge displayed a significantly higher tobramycin tolerant fraction of cells (~60%) upon the combinational treatment of AlyP1400 and tobramycin than non-treated and tobramycin-treated dispersal cells, as well as the planktonic cells (all below 10%). These results generate new knowledge about the gene expression and antibiotic resistance profiles of dispersed cells from biofilm. This information can guide the design of safer and more efficient therapeutic strategies for the combinational use of alginate lyase and tobramycin to treat P. aeruginosa biofilm-related infections in CF lungs.

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Complete Genome Sequence of Pseudomonas aeruginosa Reference Strain PAK

Abstract: We report the complete genome of Pseudomonas aeruginosa strain PAK, a strain which has been instrumental in the study of a range of P. aeruginosa virulence and pathogenesis factors and has been used for over 50 years as a laboratory reference strain.

Cited by 29 publications

References 16 publications

Pseudomonas aeruginosa is capable of natural transformation in biofilms

Pseudomonas aeruginosa is capable of natural transformation in biofilms

Pseudomonas aeruginosa is capable of natural transformation in biofilms

Characterizations of the viability and gene expression of dispersal cells from Pseudomonas aeruginosa biofilms released by alginate lyase and tobramycin

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