Single-array measurements reveal non-uniform, mosaic-like chemosensory arrays in bacteria

Abstract

Motile bacteria use supramolecular arrays to detect effector gradients in their environment. In Escherichia coli, thousands of chemoreceptor molecules with diverse sensory properties cooperatively modulate the kinase activity of these arrays and, via phosphotransfer to a diffusible response regulator, control the cells swimming behavior. Various methods have been used to study these sensory arrays in live cells, from population-level assays to single-cell measurements, revealing hierarchical coupling interactions that underlie their remarkable sensory properties. However, measuring the responses of individual arrays has remained a challenge. Here, by combining the kinase and response regulator into a functional hybrid protein that resides within the array, we directly measured the kinase responses of individual arrays in live cells. These measurements revealed highly diverse and growth-phase-dependent sensory properties of individual arrays. Even arrays within the same cell were not substantially correlated. We also observed dynamic shifts in receptor occupancy within individual arrays. Overall, these data suggest that each array contains a frozen non-uniformity, reflecting its unique assembly history and resulting in a mosaic arrangement of cooperative signaling regions, each with distinct receptor content. Consistent with this view, measured dose-responses of individual arrays mostly exhibit low cooperativity. Interestingly, signal integration in such non-uniform arrays is expected to inherently vary with the size of the cooperative regions.