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Abstract

Quorum sensing (QS) is a widespread mechanism of environment sensing and behavioural coordination in bacteria. At its core, QS is based on the production, sensing and response to small signalling molecules. Previous work with shows that QS can be used to achieve resolution and deliver a dosed response to the bacteria’s density environment, implying a sophisticated mechanism of control. To shed light on how the mechanistic signal components contribute to graded responses to density, we assess the impact of genetic (AHL signal synthase deletion) and/or signal supplementation (exogenous AHL addition) perturbations on reaction-norms to changes in density. Our approach condenses data from 2000 timeseries (over 74 000 individual observations) into a comprehensive view of QS-controlled gene expression across variation in genetic, environmental and signal determinants of expression. We first confirm that deleting either (∆) or both (∆) AHL signal synthase gene attenuates QS response to density. In the background we show persistent yet attenuated density-dependent expression due to native 3-oxo-C12-HSL signalling. We then test if density- quantities of AHL signal (3-oxo-C12-HSL, C4-HSL) added to the WT either flatten or increase responsiveness to density and find that the WT response is robust to all tested concentrations of signal, alone or in combination. We then move to progressively supplementing the genetic knockouts and find that cognate signal supplementation of a single AHL signal (∆ +3-oxo-C12-HSL, ∆ +C4HSL) is sufficient to restore the ability to respond in a density-dependent manner to increasing density. We also find that dual signal supplementation of the double AHL synthase knockout restores the ability to produce a graded response to increasing density, despite adding a density- amount of signal. Only the addition of high concentrations of both AHLs and PQS can force maximal expression and ablate responsiveness to density. Our results show that density-dependent control of expression is robust to multiple combinations of QS gene deletion and density-independent signal supplementation. Our work develops a modular approach to query the robustness and mechanistic bases of the central environmental phenotype of quorum sensing.

Funding
This study was supported by the:
  • Army Research Office (Award W911NF1910384)
    • Principle Award Recipient: SamP. Brown
  • Cystic Fibrosis Foundation (Award BROWN21P0)
    • Principle Award Recipient: SamP. Brown
  • Centers for Disease Control and Prevention Foundation (Award 75D30120C-09782)
    • Principle Award Recipient: SamP. Brown
  • Foundation for the National Institutes of Health (Award 1R21AI156817)
    • Principle Award Recipient: SamP. Brown
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2023-05-19
2024-03-29
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