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

doi:10.1177/25152564231153621

Complementation Assay Using Fusion of Split-GFP and TurboID (CsFiND) Enables Simultaneous Visualization and Proximity Labeling of Organelle Contact Sites in Yeast

Contact (Thousand Oaks). 2023 Feb 6:6:25152564231153621. doi: 10.1177/25152564231153621. eCollection 2023 Jan-Dec.

Authors

Shintaro Fujimoto¹, Shinya Tashiro¹, Yasushi Tamura¹

Affiliation

¹ Faculty of Science, Yamagata University, Yamagata, Japan.

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.

Keywords: ERMES; endoplasmic reticulum; mitochondria; split GFP; split TurboID; yeast.