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
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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.

Abstract Image

Abstract Image

Abstract Image

利用Split-GFP和TurboID (CsFiND)的融合进行互补分析,可以同时可视化和接近标记酵母细胞器接触位点。
大量研究表明,细胞器膜接触位点(MCSs)在多种细胞事件中发挥重要作用,包括脂质和离子在相连细胞器之间的运输。为了了解MCS的功能,有必要揭示MCS中积累的蛋白质。在这里,我们开发了一种名为CsFiND的互补分析系统(利用split-GFP和TurboID融合的互补分析),用于同时可视化mcs和鉴定mcs定位蛋白。我们在酵母内质网和线粒体外膜上表达了CsFiND蛋白,以验证CsFiND作为鉴定mcs定位蛋白的工具的可靠性。
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