Modular and Single-Cell Sensors of Bacterial Ser/Thr Kinase Activity.

ABSTRACT: At the single-cell level, protein kinase activity is typically inferred from downstream transcriptional reporters. However, promoters are often coregulated by several pathways, making the activity of a specific kinase difficult to deconvolve. Here, we present modular, direct, and specific sensors of bacterial kinase activity, including FRET-based sensors, as well as a synthetic transcription factor based on the lactose repressor (LacI) that has been engineered to respond to phosphorylation. We demonstrate the utility of these sensors in measuring the activity of PrkC, a conserved bacterial Ser/Thr kinase, in different growth conditions from single cells to colonies. We also show that PrkC activity increases in response to a cell-wall active antibiotic that blocks the late steps in peptidog

SUBMITTER: Zheng CR 

PROVIDER: S-EPMC8498941 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature