Journal
MBIO
Volume -, Issue -, Pages -Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/mbio.02039-23
Keywords
Pseudomonas aeruginosa; quorum sensing; gene regulation; single cell analysis; heterogeneity; LasR; RsaL
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Quorum sensing allows single cells to synchronize gene expression in bacterial populations, but studies have shown significant cell-to-cell variation in this process, resulting in coexisting subpopulations of active and inactive cells. The molecular mechanisms behind this phenomenon, particularly in the las QS system of Pseudomonas aeruginosa, have not been fully defined.
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.
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