出芽酵母のオートファゴソーム形成に关わる atg タンパク质局在の可视化 .pdfVIP

出芽酵母のオートファゴソーム形成に関わるAtg タンパク質局在の可視化 Visualization of autophagy-related proteins during autophagosome biogenesis in Saccharomyces cerevisiae （日本細胞生物学会推薦） 代表研究者 東京大学 鈴木邦律 The University of Tokyo Kuninori SUZUKI Macroautophagy (autophagy) is a highly conserved cellular recycling process involved in degradation of eukaryotic cellular components. During autophagy, macromolecules and organelles are sequestered into the double-membrane autophagosome and degraded in the vacuole/lysosome. Autophagy-related 8 (Atg8), a core Atg protein essential for autophagosome formation, is a marker of several autophagic structures: the pre-autophagosomal structure (PAS), isolation membrane (IM), and autophagosome. Atg8 is conjugated to a phospholipid phosphatidylethanolamine (PE) through a ubiquitin-like conjugation system to yield Atg8-PE; this reaction is called Atg8 lipidation. Although the mechanisms of Atg8 lipidation have been well studied in vitro, the cellular locale of Atg8 lipidation remains enigmatic. Atg3 is an E2-like enzyme that catalyzes the conjugation reaction between Atg8 and PE. Therefore, I hypothesized that the localization of Atg3 would provide insights about the site of the lipidation reaction. To explore this idea, we constructed functional GFP-tagged Atg3 (Atg3-GFP). During autophagy, Atg3-GFP transiently formed a single dot per cell on the vacuolar membrane. This Atg3-GFP dot colocalized with 2×mCherry-tagged Atg8, demonstrating that Atg3 is localized to the IM by fine-localization analysis. The localization of Atg3 suggests that Atg3 plays an important role in autophagosome formation at the IM. 研究目的 のような分子機構でオルガネラの膜構造を構築して 真核細胞の中では、細胞周期や環境条件に応じて いくのかを解明することを目的とする。 オルガネラが崩壊し、再構成される現象が頻繁に見