Designing genetically encoded probes to quantify nucleotide cofactors during the cell cycle in Caulobacter crescentus

Abstract

A recent study in Caulobacter crescentus, a dimorphic bacterial model to study cell cycle in prokaryotes, revealed the existence of a dynamic cytoplasmic redox and its importance in cell cycle progression. It was demonstrated that the cytoplasm of the swarmer (G1) cell remained reduced and a transition to oxidized state occurred during the G1 to S transition, which returned to reduced state at the G2/M phase. The importance of metabolic currencies such as NAD(P)(H) has been well documented to be involved in regulating the redox state of the cell. While NADH has been shown to cause an oxidative environment, NADPH has been demonstrated to act as a reducing equivalent for cells under oxidative stress. In Caulobacter, the status of NAD(P)(H) during the cell cycle, and redox oscillation, remains unexplored. Considering the influence of NAD(P)(H) on redox, it is likely that change in NAD(P)(H) levels could occur during the Caulobacter cell cycle. This study is focused towards designing genetically encoded sensors to measure the levels of nucleotide co-factors in live Caulobacter cells.