Mining and Quantifying In Vivo Molecular Interactions in Abiotic Stress Acclimation.

Abstract

Stress acclimation is initialized by sensing the stressor, transducing the signal, and inducing the response. In particular, the signal transduction is driven by protein-protein interactions and the response might involve de novo complex formation, shifts in subcellular localization and, thus, transportation that is mediated by other proteins. The investigation of protein-protein interactions and their regulation upon abiotic stress is crucial for a deeper understanding of the underlying mechanisms. FRET measurements by sensitized emission allow for the analysis of protein-protein interactions in real time and have a high potential to provide new insights into the regulation of protein-protein interaction with respect to subcellular localization and time. Within this section protocols are provided which allow for FRET analysis on the single cell level, the image acquisition procedure is described in detail and ImageJ plugins are suggested for the data evaluation.