Marta Mellini
RsaL-driven negative regulation promotes heterogeneity in Pseudomonas aeruginosa quorum sensing
Mellini, Marta; Letizia, Morgana; Caruso, Lorenzo; Guiducci, Alessandra; Meneghini, Carlo; Heeb, Stephan; Williams, Paul; Cámara, Miguel; Visca, Paolo; Imperi, Francesco; Leoni, Livia; Rampioni, Giordano
Authors
Morgana Letizia
Lorenzo Caruso
Alessandra Guiducci
Carlo Meneghini
Dr STEPHAN HEEB stephan.heeb@nottingham.ac.uk
Assistant Professor
PAUL WILLIAMS PAUL.WILLIAMS@NOTTINGHAM.AC.UK
Professor of Molecular Microbiology
MIGUEL CAMARA MIGUEL.CAMARA@NOTTINGHAM.AC.UK
Professor of Molecular Microbiology
Paolo Visca
Francesco Imperi
Livia Leoni
Giordano Rampioni
Contributors
Joanna B. Goldberg
Editor
Abstract
In its canonical interpretation, quorum sensing (QS) allows single cells in a bacterial population to synchronize gene expression and hence perform specific tasks collectively once the quorum cell density is reached. However, growing evidence in different bacterial species indicates that considerable cell-to-cell variation in the QS activation state occurs during growth, often resulting in coexisting subpopulations of cells in which QS is active (quorate cells) or inactive (non-quorate cells). Heterogeneity has been observed in the las QS system of the opportunistic pathogen Pseudomonas aeruginosa. However, the molecular mechanisms underlying this phenomenon have not yet been defined. The las QS system consists of an incoherent feedforward loop in which the LasR transcriptional regulator activates the expression of the lasI synthase gene and rsaL, coding for the lasI transcriptional repressor RsaL. Here, single-cell-level gene expression analyses performed in ad hoc engineered biosensor strains and deletion mutants revealed that direct binding of RsaL to the lasI promoter region increases heterogeneous activation of the las QS system. Experiments performed with a dual-fluorescence reporter system showed that the LasR-dependent expression of lasI and rsaL does not correlate in single cells, indicating that RsaL acts as a brake that stochastically limits the transition of non-quorate cells to the quorate state in a subpopulation of cells expressing high levels of this negative regulator. Interestingly, the rhl QS system that is not controlled by an analogous RsaL protein showed higher homogeneity with respect to the las system.
Citation
Mellini, M., Letizia, M., Caruso, L., Guiducci, A., Meneghini, C., Heeb, S., Williams, P., Cámara, M., Visca, P., Imperi, F., Leoni, L., & Rampioni, G. (2023). RsaL-driven negative regulation promotes heterogeneity in Pseudomonas aeruginosa quorum sensing. mBio, 14(6), Article e02039-23. https://doi.org/10.1128/mbio.02039-23
Journal Article Type | Article |
---|---|
Acceptance Date | Sep 5, 2023 |
Online Publication Date | Oct 16, 2023 |
Publication Date | Dec 19, 2023 |
Deposit Date | Oct 21, 2023 |
Publicly Available Date | Oct 26, 2023 |
Journal | mBio |
Print ISSN | 2161-2129 |
Electronic ISSN | 2150-7511 |
Publisher | American Society for Microbiology |
Peer Reviewed | Peer Reviewed |
Volume | 14 |
Issue | 6 |
Article Number | e02039-23 |
DOI | https://doi.org/10.1128/mbio.02039-23 |
Keywords | Pseudomonas aeruginosa; quorum sensing; gene regulation; single cell analysis; heterogeneity; LasR; RsaL |
Public URL | https://nottingham-repository.worktribe.com/output/26261889 |
Publisher URL | https://journals.asm.org/doi/10.1128/mbio.02039-23 |
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Publisher Licence URL
https://creativecommons.org/licenses/by/4.0/
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