Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit
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 relea...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2022
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/154009 |
| _version_ | 1811677829058789376 |
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| author | Soh, Eliza Ye-Chen Smith, Frances Gimenez, Maxime Rémi Yang, Liang Vejborg, Rebecca Munk Fletcher, Matthew Halliday, Nigel Bleves, Sophie Heeb, Stephan Cámara, Miguel Givskov, Michael Hardie, Kim R. Tolker-Nielsen, Tim Ize, Bérengère Williams, Paul |
| author2 | Singapore Centre for Environmental Life Sciences and Engineering (SCELSE) |
| author_facet | Singapore Centre for Environmental Life Sciences and Engineering (SCELSE) Soh, Eliza Ye-Chen Smith, Frances Gimenez, Maxime Rémi Yang, Liang Vejborg, Rebecca Munk Fletcher, Matthew Halliday, Nigel Bleves, Sophie Heeb, Stephan Cámara, Miguel Givskov, Michael Hardie, Kim R. Tolker-Nielsen, Tim Ize, Bérengère Williams, Paul |
| author_sort | Soh, Eliza Ye-Chen |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T02:43:35Z |
| format | Journal Article |
| id | ntu-10356/154009 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:43:35Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1540092022-08-12T06:53:12Z Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit Soh, Eliza Ye-Chen Smith, Frances Gimenez, Maxime Rémi Yang, Liang Vejborg, Rebecca Munk Fletcher, Matthew Halliday, Nigel Bleves, Sophie Heeb, Stephan Cámara, Miguel Givskov, Michael Hardie, Kim R. Tolker-Nielsen, Tim Ize, Bérengère Williams, Paul Singapore Centre for Environmental Life Sciences and Engineering (SCELSE) Science::Geology Ampicillin Antibiotic Agent 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. Published version This work was supported via grants to PW, MC and KH from the 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). BI was supported by a grant from Vaincre la Mucoviscidose and the Gre´gory Lemarchal associations (RF20140501138). TTN and MG were supported by grants from the 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. 2022-06-07T05:19:08Z 2022-06-07T05:19:08Z 2021 Journal Article Soh, E. Y., 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 bc₁ sub-unit. PLOS Pathogens, 17(8), e1009425-. https://dx.doi.org/10.1371/journal.ppat.1009425 1553-7366 https://hdl.handle.net/10356/154009 10.1371/journal.ppat.1009425 34460871 2-s2.0-85114117570 8 17 e1009425 en PLOS Pathogens © 2021 Soh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. application/pdf |
| spellingShingle | Science::Geology Ampicillin Antibiotic Agent Soh, Eliza Ye-Chen Smith, Frances Gimenez, Maxime Rémi Yang, Liang Vejborg, Rebecca Munk Fletcher, Matthew Halliday, Nigel Bleves, Sophie Heeb, Stephan Cámara, Miguel Givskov, Michael Hardie, Kim R. Tolker-Nielsen, Tim Ize, Bérengère Williams, Paul Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title | Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title_full | Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title_fullStr | Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title_full_unstemmed | Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title_short | Disruption of the Pseudomonas aeruginosa Tat system perturbs PQS-dependent quorum sensing and biofilm maturation through lack of the Rieske cytochrome bc₁ sub-unit |
| title_sort | disruption of the pseudomonas aeruginosa tat system perturbs pqs dependent quorum sensing and biofilm maturation through lack of the rieske cytochrome bc₁ sub unit |
| topic | Science::Geology Ampicillin Antibiotic Agent |
| url | https://hdl.handle.net/10356/154009 |
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