Rapamycin-dependent delocalization as a novel tool to reveal protein-protein interactions in plants [article]

Joanna Winkler, Evelien Mylle, Andreas De Meyer, Benjamin Pavie, Julie Merchie, Peter Grones, Daniel Van Damme
2020 bioRxiv   pre-print
AbstractIdentifying protein-protein interactions (PPI) is crucial to understand any type of biological process. Many PPI tools are available, yet only some function within the context of a plant cell. Narrowing down even further, only few PPI tools allow visualizing higher order interactions. Here, we present a novel and conditional in vivo PPI tool for plant research. Knocksideways in plants (KSP) uses the ability of rapamycin to alter the localization of a bait protein and its interactors via
more » ... the heterodimerization of FKBP and FRB domains. KSP is inherently free from many limitations, which other PPI systems hold. It is an in vivo tool, it is flexible concerning the orientation of protein tagging as long as this does not interfere with the interaction and it is compatible with a broad range of fluorophores. KSP is also a conditional tool and therefore does not require additional controls. The interactions can be quantified and in high throughput by the scripts that we provide. Finally, we demonstrate that KSP can visualize higher-order interactions. It is therefore a versatile tool, complementing the PPI methods field with unique characteristics and applications.
doi:10.1101/2020.03.09.983270 fatcat:kpqmvkc6lzbq3mu5q45jlx3jhu