Live cell imaging of redox mediated nrf2 activity using transcription factor based biosensors and chemogenetic tools

Official URL: https://risc01.sabanciuniv.edu/record=b2766815_(Table of contents)

Nuclear factor erythroid 2-related factor 2 (Nrf2) controls the expression of multiple antioxidant response element dependent genes to regulate the (patho)physiological oxidant mediated responses. Auranofin is a gold(I) containing drug of which the pharmacological activity is associated with its ability to induce Nrf2 activation. However, the mechanism of auranofin-mediated Nrf2 activation remains unclear. Auranofin also increases ROS production in the cell. We investigated the role of hydrogen peroxide (H2O2) on auranofin-mediated Nrf2 activation. In this study, we exploited chemogenetic D-amino acid oxidase (DAAO) approaches that permit the generation of H2O2 in single cells with high spatio-temporal resolution. we employed transcription factor-based biosensor called pLG4.mCherryNrf2 to visualize Nrf2 activation. By using genetically encoded biosensor HyPer7, we monitored H2O2 in different subcellular locales. Our findings showed that auranofin induces intracellular production of H2O2 while triggering Nrf2 activity, however, bolus administration of exogenous H2O2 or chemogenetically induced endogenous H2O2 generation did not change Nrf2 activation. Our findings draw new picture of oxidative stress dependent Nrf2 activation in cultured cells.