Visualizing the Perturbation
of Cellular Cyclic di-GMP
Levels in Bacterial Cells
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Abstract
Cyclic di-GMP (c-di-GMP) has emerged as a prominent intracellular
messenger that coordinates biofilm formation and pathogenicity in
many bacterial species. Developing genetically encoded biosensors
for c-di-GMP will help us understand how bacterial cells respond to
environmental changes via the modulation of cellular c-di-GMP levels.
Here we report the design of two genetically encoded c-di-GMP fluorescent
biosensors with complementary dynamic ranges. By using the biosensors,
we found that several compounds known to promote biofilm dispersal
trigger a decline in c-di-GMP levels in Escherichia
coli cells. In contrast, cellular c-di-GMP levels
were elevated when the bacterial cells were treated with subinhibitory
concentrations of biofilm-promoting antibiotics. The biosensors also
revealed that E. coli cells engulfed
by macrophages exhibit lower c-di-GMP levels, most likely as a response
to the enormous pressures of survival during phagocytosis