Research Projects

Escherichia coli cells, like those in the human gut, face various stressful conditions, such as nutrient deprivation, osmolarity changes, low pH, and oxidants. These environments lead to the development of adaptive stress responses, including well-orchestrated signaling cascades and gene regulation at the transcriptional level. Variable DNA supercoiling is crucial for controlling gene regulation in bacteria, and inhibition of DNA gyrase can alter its levels. Environmental stresses, such as nutritional, acid, oxidative, osmotic, and thermal stresses, can alter the extent of supercoiling. YacG, a gyrase modulatory protein from E. coli, employs a bipartite mechanism to inhibit enzyme activity. Although YacG-DNA gyrase interaction has been well characterized in vitro, its biological significance remains elusive. The project proposal aims to unravel the physiological basis for YacG-mediated increased tolerance to Fluoroquinolones. The impact of YacG expression on E. coli cell viability will be assessed under different stress conditions, including Fluoroquinolone-induced stress. In vivo supercoiling levels and gyrase expression will be monitored under stress conditions, and the correlation between YacG levels and genome supercoiling status will be probed. Coimmunoprecipitation studies will be performed to assess Gyrase-YacG interaction under stress conditions. Insights gained from this study could help develop improved combinatorial therapeutic strategies to counter antimicrobial resistance.