Complementation Assay Using Fusion of Split-GFP and TurboID (CsFiND) Enables Simultaneous Visualization and Proximity Labeling of Organelle Contact Sites in Yeast
Shintaro Fujimoto, Shinya Tashiro, Yasushi Tamura
Abstract
Numerous studies have revealed that organelle membrane contact sites (MCSs) play important roles in diverse cellular events, including the transport of lipids and ions between connected organelles. To understand MCS functions, it is essential to uncover proteins that accumulate at MCSs. Here, we develop a complementation assay system termed CsFiND (Complementation assay using Fusion of split-GFP and TurboID) for the simultaneous visualization of MCSs and identification of MCS-localized proteins. We express the CsFiND proteins on the endoplasmic reticulum and mitochondrial outer membrane in yeast to verify the reliability of CsFiND as a tool for identifying MCS-localized proteins.
Topics & Concepts
ComplementationOrganelleEndoplasmic reticulumYeastBimolecular fluorescence complementationCell biologyGreen fluorescent proteinProtein-fragment complementation assayCytoplasmBiologyFusion proteinChemistryBiochemistryGenePhenotypeRecombinant DNAMitochondrial Function and PathologyCellular transport and secretionEndoplasmic Reticulum Stress and Disease