Pseudomonas aeruginosa Biofilm, a Programmed Bacterial Life for Fitness

Lee, Keehoon; Yoon, Sang Sun

doi:10.4014/jmb.1611.11056

Cited by 169 publications

(114 citation statements)

References 115 publications

(193 reference statements)

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“…The data presented here suggest that the intraisolate variability of these factors alters OMV production, which may play an important part in bacterial fitness in natural environments and within the host. P. aeruginosa is known to grow in a biofilm both in the environment and in infectious niches, such as the airways of patients with cystic fibrosis (39,40). High levels of extracellular DNA (eDNA) production have been linked to biofilm development and propagation (41,42).…”

Section: Figmentioning

confidence: 99%

Genomic and Phenotypic Diversity among Ten Laboratory Isolates of Pseudomonas aeruginosa PAO1

et al. 2019

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Pseudomonas aeruginosa is an opportunistic pathogen found ubiquitously in the environment and commonly associated with airway infection in patients with cystic fibrosis. P. aeruginosa strain PAO1 is one of the most commonly used laboratory-adapted research strains and is a standard laboratory-adapted strain in multiple laboratories and strain banks worldwide. Due to potential isolate-to-isolate variability, we investigated the genomic and phenotypic diversity among 10 PAO1 strains (henceforth called sublines) obtained from multiple research laboratories and commercial sources. Genomic analysis predicted a total of 5,682 genes, with 5,434 (95.63%) being identical across all 10 strains. Phenotypic analyses revealed comparable growth phenotypes in rich media and biofilm formation profiles. Limited differences were observed in antibiotic susceptibility profiles and immunostimulatory potential, measured using heat-killed whole-cell preparations in four immortalized cell lines followed by quantification of interleukin-6 (IL-6) and IL-1β secretion. However, variability was observed in the profiles of secreted molecular products, most notably, in rhamnolipid, pyoverdine, pyocyanin, Pseudomonas quinolone signal (PQS), extracellular DNA, exopolysaccharide, and outer membrane vesicle production. Many of the observed phenotypic differences did not correlate with subline-specific genetic changes, suggesting alterations in transcriptional and translational regulation. Taken together, these results suggest that individually maintained sublines of PAO1, even when acquired from the same parent subline, are continuously undergoing microevolution during culture and storage that results in alterations in phenotype, potentially affecting the outcomes of in vitro phenotypic analyses and in vivo pathogenesis studies. IMPORTANCE Laboratory-adapted strains of bacteria are used throughout the world for microbiology research. These prototype strains help keep research data consistent and comparable between laboratories. However, we have observed phenotypic variability when using different strains of Pseudomonas aeruginosa PAO1, one of the major laboratory-adopted research strains. Here, we describe the genomic and phenotypic differences among 10 PAO1 strains acquired from independent sources over 15 years to understand how individual maintenance affects strain characteristics. We observed limited genomic changes but variable phenotypic changes, which may have consequences for cross-comparison of data generated using different PAO1 strains. Our research highlights the importance of limiting practices that may promote the microevolution of model strains and calls for researchers to specify the strain origin to ensure reproducibility.

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Section: Figmentioning

confidence: 99%

Genomic and Phenotypic Diversity among Ten Laboratory Isolates of Pseudomonas aeruginosa PAO1

et al. 2019

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“…The involvement of P. aeruginosa QS systems in biofilm life cycle was firstly reported by Davis et al as they demonstrated that lasI mutants form only flat and undifferentiated biofilm structures [126]. Therefore, Las system is mainly in use for mature biofilm development [127]. Other investigators reported that rhlI gene is involved in the formation of a mushroom-like structures and are also crucial in the dispersal stage controlling the production of rhamnolipids [128,129].…”

Section: Qs Systems Of P Aeruginosamentioning

confidence: 94%

Complex Signaling Networks Controlling Dynamic Molecular Changes in Pseudomonas aeruginosa Biofilm

Guła

Dorotkiewicz-Jach

Korzekwa

et al. 2019

CMC

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The environment exerts strong influence on microbes. Adaptation of microbes to changing conditions is a dynamic process regulated by complex networks. Pseudomonas aeruginosa is a life-threating, versatile opportunistic and multi drug resistant pathogen that provides a model to investigate adaptation mechanisms to environmental changes. The ability of P. aeruginosa to form biofilms and to modify virulence in response to environmental changes are coordinated by various mechanisms including two-component systems (TCS), and secondary messengers involved in quorum sensing (QS) and c-di-GMP networks (diguanylate cyclase systems, DGC). In this review, we focus on the role of c-di-GMP during biofilm formation. We describe TCS and QS signal cascades regulated by c-di-GMP in response to changes in the external environment. We present a complex signaling network dynamically changing during the transition of P. aeruginosa from the free-living to sessile mode of growth.

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“…The biofilm lifecycle is composed of several commonly reported steps 1,[6][7][8] . Initially, planktonic bacteria propel themselves to a proximal surface, followed by an irreversible attachment to the surface.…”

Section: Mainmentioning

confidence: 99%

“…We decided to introduce modifications mainly in the D3 domain of flagellin, facing outwards in the cylindrical structure of flagellum, thus minimizing possible interference with filament assembly 32 . We modulated the P. aeruginosa flagellum that is composed of 41 subunits (based on PDB accession number 5wk5) with (1) incorporated at flagellin's th position (Fig. 2a).…”

Section: Uaasmentioning

confidence: 99%

An inside look at a biofilm: Pseudomonas aeruginosa flagella bio-tracking

Ozer

Yaniv

Chetrit

et al. 2020

Preprint

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The opportunistic pathogen, Pseudomonas aeruginosa, a flagellated bacterium, is one of the top model organisms for studying biofilm formation. In order to elucidate the role of the bacteria flagella in biofilm formation, we developed a new tool for flagella bio-tracking. We have site-specifically labeled the bacterial flagella by incorporating an unnatural amino acid into the flagella monomer via genetic code expansion. This enabled us to label and track the bacterial flagella during biofilm maturation. Direct, live imaging revealed for the first-time presence and synthesis of flagella throughout the biofilm lifecycle. To ascertain the possible role of the flagella in the strength of a biofilm we produced a “flagella knockout” strain and compared its biofilm to that of the wild type strain. Results showed a one order of magnitude stronger biofilm structure in the wild type in comparison to the flagella knockout strain. This suggests a newly discovered structural role for bacterial flagella in biofilm structure, possibly acting as a scaffold. Based on our findings we suggest a new model for biofilm maturation dynamic and underscore the importance of direct evidence from within the biofilm.

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Pseudomonas aeruginosa Biofilm, a Programmed Bacterial Life for Fitness

Cited by 169 publications

References 115 publications

Genomic and Phenotypic Diversity among Ten Laboratory Isolates of Pseudomonas aeruginosa PAO1

Genomic and Phenotypic Diversity among Ten Laboratory Isolates of Pseudomonas aeruginosa PAO1

Complex Signaling Networks Controlling Dynamic Molecular Changes in Pseudomonas aeruginosa Biofilm

An inside look at a biofilm: Pseudomonas aeruginosa flagella bio-tracking

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