Significance
Microbial cells organized on solid surfaces are the most ancient form of biological communities. Yet how single cells interact with surfaces and integrate a variety of signals to establish a sessile lifestyle is poorly understood. We developed and used sensitive biosensors to determine the kinetics of second messengers’ responses to surface attachment. This allowed us to examine cell-by-cell variability of the initial signaling events and establish that some of these events depend on flagellar motor function while others do not. Environmentally determined factors, like the energetic status of the cell, can modulate all signaling events. The complex interplay between the surface interaction inputs and external conditions can now be studied using our system.
Abstract
Bacterial cells interact with solid surfaces and change their lifestyle from single free-swimming cells to sessile communal structures (biofilms). Cyclic di-guanosine monophosphate (c-di-GMP) is central to this process, yet we lack tools for direct dynamic visualization of c-di-GMP in single cells. Here, we developed a fluorescent protein–based c-di-GMP–sensing system for Escherichia coli that allowed us to visualize initial signaling events and assess the role played by the flagellar motor. The sensor was pH sensitive, and the events that appeared on a seconds’ timescale were alkaline spikes in the intracellular pH. These spikes were not apparent when signals from different cells were averaged. Instead, a signal appeared on a minutes’ timescale that proved to be due to an increase in intracellular c-di-GMP. This increase, but not the alkaline spikes, depended upon a functional flagellar motor. The kinetics and the amplitude of both the pH and c-di-GMP responses displayed cell-to-cell variability indicative of the distinct ways the cells approached and interacted with the surface. The energetic status of a cell can modulate these events. In particular, the alkaline spikes displayed an oscillatory behavior and the c-di-GMP increase was modest in the presence of glucose.
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February 08, 2022 at 03:37AM
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Signaling events that occur when cells of Escherichia coli encounter a glass surface - pnas.org
"occur" - Google News
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