Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N

The Pseudomonas aeruginosa quorum sensing (QS) network plays a key role in the adaptation to environmental changes and the control of virulence factor production in this opportunistic human pathogen. Three interlinked QS systems, namely las, rhl, and pqs, are central to the production of pyocyanin,...

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Main Authors: Higgins, Steven, Heeb, Stephan, Rampioni, Giordano, Fletcher, Mathew P., Williams, Paul, Cámara, Miguel
Format: Article
Published: Frontiers 2018
Online Access:https://eprints.nottingham.ac.uk/53091/
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author Higgins, Steven
Heeb, Stephan
Rampioni, Giordano
Fletcher, Mathew P.
Williams, Paul
Cámara, Miguel
author_facet Higgins, Steven
Heeb, Stephan
Rampioni, Giordano
Fletcher, Mathew P.
Williams, Paul
Cámara, Miguel
author_sort Higgins, Steven
building Nottingham Research Data Repository
collection Online Access
description The Pseudomonas aeruginosa quorum sensing (QS) network plays a key role in the adaptation to environmental changes and the control of virulence factor production in this opportunistic human pathogen. Three interlinked QS systems, namely las, rhl, and pqs, are central to the production of pyocyanin, a phenazine virulence factor which is typically used as phenotypic marker for analysing QS. Pyocyanin production in P. aeruginosa is a complex process involving two almost identical operons termed phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), which drive the production of phenazine-1-carboxylic acid (PCA) which is further converted to pyocyanin by two modifying enzymes PhzM and PhzS. Due to the high sequence conservation between the phz1 and phz2 operons (nucleotide identity > 98%), analysis of their individual expression by RNA hybridization, qRT-PCR or transcriptomics is challenging. To overcome this difficulty, we utilized luminescence based promoter fusions of each phenazine operon to measure in planktonic cultures their transcriptional activity in P. aeruginosa PAO1-N genetic backgrounds impaired in different components of the las, rhl, and pqs QS systems, in the presence or absence of different QS signal molecules. Using this approach, we found that all three QS systems play a role in differentially regulating the phz1 and phz2 phenazine operons, thus uncovering a higher level of complexity to the QS regulation of PCA biosynthesis in P. aeruginosa than previously appreciated.
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spelling nottingham-530912020-05-04T19:47:07Z https://eprints.nottingham.ac.uk/53091/ Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N Higgins, Steven Heeb, Stephan Rampioni, Giordano Fletcher, Mathew P. Williams, Paul Cámara, Miguel The Pseudomonas aeruginosa quorum sensing (QS) network plays a key role in the adaptation to environmental changes and the control of virulence factor production in this opportunistic human pathogen. Three interlinked QS systems, namely las, rhl, and pqs, are central to the production of pyocyanin, a phenazine virulence factor which is typically used as phenotypic marker for analysing QS. Pyocyanin production in P. aeruginosa is a complex process involving two almost identical operons termed phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), which drive the production of phenazine-1-carboxylic acid (PCA) which is further converted to pyocyanin by two modifying enzymes PhzM and PhzS. Due to the high sequence conservation between the phz1 and phz2 operons (nucleotide identity > 98%), analysis of their individual expression by RNA hybridization, qRT-PCR or transcriptomics is challenging. To overcome this difficulty, we utilized luminescence based promoter fusions of each phenazine operon to measure in planktonic cultures their transcriptional activity in P. aeruginosa PAO1-N genetic backgrounds impaired in different components of the las, rhl, and pqs QS systems, in the presence or absence of different QS signal molecules. Using this approach, we found that all three QS systems play a role in differentially regulating the phz1 and phz2 phenazine operons, thus uncovering a higher level of complexity to the QS regulation of PCA biosynthesis in P. aeruginosa than previously appreciated. Frontiers 2018-07-23 Article PeerReviewed Higgins, Steven, Heeb, Stephan, Rampioni, Giordano, Fletcher, Mathew P., Williams, Paul and Cámara, Miguel (2018) Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N. Frontiers in Cellular and Infection Microbiology, 8 . 252/1-252/13. ISSN 2235-2988 http://dx.doi.org/10.3389/fcimb.2018.00252 doi:10.3389/fcimb.2018.00252 doi:10.3389/fcimb.2018.00252
spellingShingle Higgins, Steven
Heeb, Stephan
Rampioni, Giordano
Fletcher, Mathew P.
Williams, Paul
Cámara, Miguel
Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title_full Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title_fullStr Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title_full_unstemmed Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title_short Differential regulation of the phenazine biosynthetic operons by quorum sensing in Pseudomonas aeruginosa PAO1-N
title_sort differential regulation of the phenazine biosynthetic operons by quorum sensing in pseudomonas aeruginosa pao1-n
url https://eprints.nottingham.ac.uk/53091/
https://eprints.nottingham.ac.uk/53091/
https://eprints.nottingham.ac.uk/53091/