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Quorum sensing is a widespread process in bacteria that controls collective behaviours in response to cell density. Populations of cells coordinate gene expression through the perception of self-produced chemical signals. Although this process is well-characterized genetically and biochemically, quantitative information about network properties, including induction dynamics and steady-state behaviour, is scarce. Here we integrate experiments with mathematical modelling to quantitatively analyse the LasI/LasR quorum sensing pathway in the opportunistic pathogen Pseudomonas aeruginosa . We determine key kinetic parameters of the pathway and, using the parametrized model, show that quorum sensing behaves as a bistable hysteretic switch, with stable on and off states. We investigate the significance of feedback architecture and find that positive feedback on signal production is critical for induction dynamics and bistability, whereas positive feedback on receptor expression and negative feedback on signal production play a minor role. Taken together, our data-based modelling approach reveals fundamental and emergent properties of a bacterial quorum sensing circuit, and provides evidence that native quorum sensing can indeed function as the gene expression switch it is commonly perceived to be.more » « lessFree, publicly-accessible full text available March 1, 2024
Quorum sensing is described as a widespread cell density-dependent signaling mechanism in bacteria. Groups of cells coordinate gene expression by secreting and responding to diffusible signal molecules. Theory, however, predicts that individual cells may short-circuit this mechanism by directly responding to the signals they produce irrespective of cell density. In this study, we characterize this self-sensing effect in the acyl-homoserine lactone quorum sensing system of Pseudomonas aeruginosa . We show that antiactivators, a set of proteins known to affect signal sensitivity, function to prevent self-sensing. Measuring quorum-sensing gene expression in individual cells at very low densities, we find that successive deletion of antiactivator genes qteE and qslA produces a bimodal response pattern, in which increasing proportions of constitutively induced cells coexist with uninduced cells. Comparing responses of signal-proficient and -deficient cells in cocultures, we find that signal-proficient cells show a much higher response in the antiactivator mutant background but not in the wild-type background. Our results experimentally demonstrate the antiactivator-dependent transition from group- to self-sensing in the quorum-sensing circuitry of P. aeruginosa . Taken together, these findings extend our understanding of the functional capacity of quorum sensing. They highlight the functional significance of antiactivators in the maintenance of group-level signaling and experimentally prove long-standing theoretical predictions.more » « less
Abstract A novel Compton Scanner setup has been built, commissioned and operated at the Max-Planck-Institute for Physics in Munich to collect pulses from bulk events in high-purity germanium detectors for pulse shape studies. In this fully automated setup, the detector under test is irradiated from the top with 661.660 keV gammas, some of which Compton scatter inside the detector. The interaction points in the detector can be reconstructed when the scattered gammas are detected with a pixelated camera placed at the side of the detector. The wide range of accepted Compton angles results in shorter measurement times in comparison to similar setups where only perpendicularly scattered gammas are selected by slit collimators. In this paper, the construction of the Compton Scanner, its alignment and the procedure to reconstruct interaction points in the germanium detector are described in detail. The creation of a first pulse shape library for an n-type segmented point-contact germanium detector is described. The spatial reconstruction along the beam axis is validated by a comparison to measured surface pulses. A first comparison of Compton Scanner pulses to simulated pulses is presented to demonstrate the power of the Compton Scanner to test simulation inputs and models.more » « less