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Atg15 in Saccharomyces cerevisiae consists of two functionally distinct domains
Molecular Biology of the Cell ( IF 3.1 ) Pub Date : 2021-02-24 , DOI: 10.1091/mbc.e20-07-0500
Eri Hirata 1 , Kyo Shirai 1 , Tatsuya Kawaoka 1 , Kosuke Sato 1 , Fumito Kodama 1 , Kuninori Suzuki 1, 2, 3
Affiliation  

Autophagy is a cellular degradation system widely conserved among eukaryotes. During autophagy, cytoplasmic materials fated for degradation are compartmentalized in double membrane–bound organelles called autophagosomes. After fusing with the vacuole, their inner membrane–bound structures are released into the vacuolar lumen to become autophagic bodies and eventually degraded by vacuolar hydrolases. Atg15 is a lipase essential for disintegration of autophagic body membranes and has a transmembrane domain at the N-terminus and a lipase domain at the C-terminus. However, the roles of both domains in vivo are not well understood. In this study, we found that the N-terminal domain alone can travel to the vacuole via the multivesicular body pathway, and that targeting of the C-terminal lipase domain to the vacuole is required for degradation of autophagic bodies. Moreover, we found that the C-terminal domain could disintegrate autophagic bodies when it was transported to the vacuole via the Pho8 pathway instead of the multivesicular body pathway. Finally, we identified H435 as one of the residues composing the putative catalytic triad, and W466 as an important residue for degradation of autophagic bodies. This study may provide a clue to understanding how the C-terminal lipase domain recognizes autophagic bodies to degrade them.

Video S1: Cells expressing C-terminally GFP-fused Atg15 were grown in SDCA medium to mid-log phase and treated with rapamycin for 2 h before observation. All videos are played at 0.8 times speed. (A) Wild-type cells. (B) pep4Δ cells.Download Original Video (.1 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e20-07-0500/20210222/media/mc-e20-07-0500-s08.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab3a0b97e9f37f414d52fd2cdf7c979b94bfe423ab53601896b21c24040801869b86c42fdee41116cf373243fb61ee7faf14621622b1c5725f0db77068efb741538a6b7b5e0d324d695d811cd0e21499a75bc40Video S1: Cells expressing C-terminally GFP-fused Atg15 were grown in SDCA medium to mid-log phase and treated with rapamycin for 2 h before observation. All videos are played at 0.8 times speed. (A) Wild-type cells. (B) pep4Δ cells.Download Original Video (.1 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e20-07-0500/20210222/media/mc-e20-07-0500-s09.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab3a0b97e9f37f414d52fd2cdf7c979b94bfe423ab53601896b21c24040801869b86c42fdee41116cf373253fb61ee7faf1460d88cdc6e908dc8db7b98e054285a4f1bb6d16867307e1e8f4796638c3d68f7dfb


中文翻译:

酿酒酵母中的 Atg15 由两个功能不同的域组成

自噬是一种在真核生物中广泛保守的细胞降解系统。在自噬过程中,注定要降解的细胞质材料被分隔在称为自噬体的双膜结合细胞器中。与液泡融合后,它们的内膜结合结构被释放到液泡腔内成为自噬体,最终被液泡水解酶降解。Atg15 是自噬体膜分解所必需的脂肪酶,在 N 端具有跨膜结构域,在 C 端具有脂肪酶结构域。然而,这两个结构域在体内的作用不太了解。在这项研究中,我们发现单独的 N 端结构域可以通过多泡体途径到达液泡,并且自噬体降解需要将 C 端脂肪酶结构域靶向液泡。此外,我们发现当 C 端结构域通过 Pho8 途径而不是多泡体途径转运到液泡时,它可以分解自噬体。最后,我们确定 H435 是构成推定催化三联体的残基之一,W466 是降解自噬体的重要残基。这项研究可能为理解 C 端脂肪酶结构域如何识别自噬体以降解它们提供线索。

视频 S1:表达 C 端 GFP 融合 Atg15 的细胞在 SDCA 培养基中生长至对数中期,并在观察前用雷帕霉素处理 2 小时。所有视频都以 0.8 倍的速度播放。(A) 野生型细胞。(B) pep4Δ 细胞。下载原始视频 (.1 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e20-07-0500/20210222/media/mc-e20-07-0500-s08.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab3a0b97e9f37f414d52fd2cdf7c979b94bfe423ab53601896b21c24040801869b86c42fdee41116cf373243fb61ee7faf14621622b1c5725f0db77068efb741538a6b7b5e0d324d695d811cd0e21499a75bc40视频 S1:表达 C 端 GFP 融合 Atg15 的细胞在 SDCA 培养基中生长至对数中期,并在观察前用雷帕霉素处理 2 小时。所有视频都以 0.8 倍的速度播放。(A) 野生型细胞。(B) pep4Δ 细胞。下载原始视频 (.1 MB)https://ascb-prod-streaming.literatumonline.com/journals/content/mboc/0/mboc.ahead-of-print/mbc.e20-07-0500/20210222/media/mc-e20-07-0500-s09.,652,642,.mp4.m3u8?b92b4ad1b4f274c70877518515abb28bda92fbabe7b929571bd415190bf44d1790b7324fb90f184f91fab10e395327a1bef9725d0c99a358bbbe69c082a95f9d49957eb2932b2be9a6f328f5ec3ab3a0b97e9f37f414d52fd2cdf7c979b94bfe423ab53601896b21c24040801869b86c42fdee41116cf373253fb61ee7faf1460d88cdc6e908dc8db7b98e054285a4f1bb6d16867307e1e8f4796638c3d68f7dfb
更新日期:2021-02-24
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