Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc1sub-unit
| Autor: | Maxime Rémi Gimenez, Liang Yang, Miguel Cámara, Tim Tolker-Nielsen, Frances R. Smith, Stephan Heeb, Eliza Ye-Chen Soh, Rebecca Munk Vejborg, Paul Williams, Bérengère Ize, Matthew P. Fletcher, Sophie Bleves, Kim R. Hardie, Michael Givskov, Nigel Halliday |
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| Přispěvatelé: | University of Nottingham, UK (UON), Laboratoire d'ingénierie des systèmes macromoléculaires (LISM), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Nanyang Technological University [Singapour], University of Copenhagen = Københavns Universitet (UCPH), Aix Marseille Université (AMU), Biotechnology and Biological Sciences Research Council (BBSRC), U.K. (BB/F014392/1), BBSRC/National Biofilms Innovation Centre (BB/R012415/1), the Medical Research Council U.K. (MR/N501852/1), the Wellcome Trust (103884/Z/14/Z and 108876/Z/15/Z) and the European Union FP7 collaborative action grant (NABATIVI, 223670), Vaincre la Mucoviscidose and the Grégory Lemarchal associations (RF20140501138), Danish Strategic Research Council (9040-00023B), the Danish Council for Independent Research (09-065732), the Novo Nordisk Foundation (Biofilm2-AEEE/CPH) and the Lundbeck Foundation (2015‐486), Denmark, IZE, BERENGERE, Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), University of Copenhagen = Københavns Universitet (KU) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Mutant
Quinolones Pathology and Laboratory Medicine [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology medicine.disease_cause Biochemistry Electron Transport Complex III chemistry.chemical_compound Antibiotics Medicine and Health Sciences Confocal laser microscopy Biology (General) [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Twin-Arginine-Translocation System 0303 health sciences Antimicrobials Effector Quorum Sensing Drugs Anti-Bacterial Agents Bacterial Pathogens Extracellular Matrix Polymerase chain reaction Cell biology Mutant Strains Phenotypes Medical Microbiology Pseudomonas aeruginosa Pathogens Cellular Structures and Organelles Research Article DNA Bacterial Virulence Factors QH301-705.5 Immunology Research and Analysis Methods Biosynthesis Microbiology Extracellular Vesicles 03 medical and health sciences Pyocyanin Bacterial Proteins Pseudomonas Microbial Control Virology Genetics medicine [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Molecular Biology Techniques Microbial Pathogens Molecular Biology 030304 developmental biology Pharmacology Molecular Biology Assays and Analysis Techniques Bacteria 030306 microbiology Organisms Wild type Biofilm Biology and Life Sciences Bacteriology [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology Gene Expression Regulation Bacterial Cell Biology RC581-607 [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology Quorum sensing Bacterial biofilms chemistry Biofilms Coenzyme Q – cytochrome c reductase Mutation Pyocyanine Library screening Parasitology Gene expression Glycolipids [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology Immunologic diseases. Allergy |
| Zdroj: | Soh, E Y C, Smith, F, Gimenez, M R, Yang, L, Vejborg, R M, Fletcher, M, Halliday, N, Bleves, S, Heeb, S, Cámara, M, Givskov, M, Hardie, K R, Tolker-Nielsen, T, Ize, B & Williams, P 2021, ' Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc 1 sub-unit ', PLOS Pathogens, vol. 17, no. 8, e1009425 . https://doi.org/10.1371/journal.ppat.1009425 PLoS Pathogens, Vol 17, Iss 8, p e1009425 (2021) PLoS Pathogens PLoS Pathogens, 2021, 17 (8), pp.e1009425. ⟨10.1371/journal.ppat.1009425⟩ PLoS Pathogens, Public Library of Science, 2021, 17 (8), pp.e1009425. ⟨10.1371/journal.ppat.1009425⟩ |
| ISSN: | 1553-7366 1553-7374 |
| Popis: | Extracellular DNA (eDNA) is a major constituent of the extracellular matrix of Pseudomonas aeruginosa biofilms and its release is regulated via pseudomonas quinolone signal (PQS) dependent quorum sensing (QS). By screening a P. aeruginosa transposon library to identify factors required for DNA release, mutants with insertions in the twin-arginine translocation (Tat) pathway were identified as exhibiting reduced eDNA release, and defective biofilm architecture with enhanced susceptibility to tobramycin. P. aeruginosa tat mutants showed substantial reductions in pyocyanin, rhamnolipid and membrane vesicle (MV) production consistent with perturbation of PQS-dependent QS as demonstrated by changes in pqsA expression and 2-alkyl-4-quinolone (AQ) production. Provision of exogenous PQS to the tat mutants did not return pqsA, rhlA or phzA1 expression or pyocyanin production to wild type levels. However, transformation of the tat mutants with the AQ-independent pqs effector pqsE restored phzA1 expression and pyocyanin production. Since mutation or inhibition of Tat prevented PQS-driven auto-induction, we sought to identify the Tat substrate(s) responsible. A pqsA::lux fusion was introduced into each of 34 validated P. aeruginosa Tat substrate deletion mutants. Analysis of each mutant for reduced bioluminescence revealed that the primary signalling defect was associated with the Rieske iron-sulfur subunit of the cytochrome bc1 complex. In common with the parent strain, a Rieske mutant exhibited defective PQS signalling, AQ production, rhlA expression and eDNA release that could be restored by genetic complementation. This defect was also phenocopied by deletion of cytB or cytC1. Thus, either lack of the Rieske sub-unit or mutation of cytochrome bc1 genes results in the perturbation of PQS-dependent autoinduction resulting in eDNA deficient biofilms, reduced antibiotic tolerance and compromised virulence factor production. Author summary Pseudomonas aeruginosa is a highly adaptable human pathogen responsible for causing chronic biofilm-associated infections. Biofilms are highly refractory to host defences and antibiotics and thus difficult to eradicate. The biofilm extracellular matrix incorporates extracellular DNA (eDNA). This stabilizes biofilm architecture and helps confer tolerance to antibiotics. Since mechanisms that control eDNA release are not well understood, we screened a P. aeruginosa mutant bank for strains with defects in eDNA release and discovered a role for the twin-arginine translocation (Tat) pathway that exports folded proteins across the cytoplasmic membrane. Perturbation of the Tat pathway resulted in defective biofilms susceptible to antibiotic treatment as a consequence of perturbed pseudomonas quinolone (PQS) signalling. This resulted in the failure to produce or release biofilm components including eDNA, phenazines and rhamnolipids as well as microvesicles. Furthermore, we discovered that perturbation of PQS signalling was a consequence of the inability of tat mutants to translocate the Rieske subunit of the cytochrome bc1 complex involved in electron transfer and energy transduction. Given the importance of PQS signalling and the Tat system to virulence and biofilm maturation in P. aeruginosa, our findings underline the potential of the Tat system as a drug target for novel antimicrobial agents. |
| Databáze: | OpenAIRE |
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