エンドウ トシヤ   ENDO TOSHIYA
  遠藤 斗志也
   所属   京都産業大学  生命科学部 先端生命科学科
   職種   客員教授
言語種別 英語
発行・発表の年月 2018/12/01
形態種別 研究論文
査読 査読あり
標題 Visualizing multiple inter-organelle contact sites using the organelle-Targeted split-GFP system
執筆形態 その他
掲載誌名 Scientific Reports
出版社・発行元 Nature Publishing Group
巻・号・頁 8(1),pp.6175
著者・共著者 Yuriko Kakimoto,Shinya Tashiro,Rieko Kojima,Yuki Morozumi,Toshiya Endo,Yasushi Tamura
概要 Functional integrity of eukaryotic organelles relies on direct physical contacts between distinct organelles. However, the entity of organelle-Tethering factors is not well understood due to lack of means to analyze inter-organelle interactions in living cells. Here we evaluate the split-GFP system for visualizing organelle contact sites in vivo and show its advantages and disadvantages. We observed punctate GFP signals from the split-GFP fragments targeted to any pairs of organelles among the ER, mitochondria, peroxisomes, vacuole and lipid droplets in yeast cells, which suggests that these organelles form contact sites with multiple organelles simultaneously although it is difficult to rule out the possibilities that these organelle contacts sites are artificially formed by the irreversible associations of the split-GFP probes. Importantly, split-GFP signals in the overlapped regions of the ER and mitochondria were mainly co-localized with ERMES, an authentic ER-mitochondria tethering structure, suggesting that split-GFP assembly depends on the preexisting inter-organelle contact sites. We also confirmed that the split-GFP system can be applied to detection of the ER-mitochondria contact sites in HeLa cells. We thus propose that the split-GFP system is a potential tool to observe and analyze inter-organelle contact sites in living yeast and mammalian cells.
DOI 10.1038/s41598-018-24466-0
ISSN 2045-2322
PMID 29670150