An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure.
An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure.
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DOI：
10.1038/s41467-018-05254-w
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发表时间：
2018-07-20
影响因子：
16.6
通讯作者：
Wang HG
中科院分区：
文献类型：
--
作者：
Takahashi Y;He H;Tang Z;Hattori T;Liu Y;Young MM;Serfass JM;Chen L;Gebru M;Chen C;Wills CA;Atkinson JM;Chen H;Abraham T;Wang HG
由于区分非闭合和闭合自噬体膜的技术限制，吞噬体闭合的机制尚不清楚。在这里，我们报道了HaloTag-LC3自噬体完成实验，该实验可以特异性检测吞噬细胞、新生自噬体和成熟的自噬结构。通过该试验，我们确定了运输所需的内体分选复合物(ESCRT)-III组分CHMP2A是吞噬体关闭的关键调节剂。在自噬过程中，CHMP2A易位到吞噬细胞，调节自噬体内外膜的分离，形成双膜自噬体。一致地，抑制aaa - atp酶VPS4活性会损害自噬体的完成。escrt介导的膜脱落似乎是通过防止溶酶体相关膜糖蛋白1错定位到内自噬体膜而形成功能性自噬体的关键步骤。总之，我们的工作揭示了ESCRT机制在自噬体形成的最后一步中的作用，并为自噬体生物发生和成熟的定量分析提供了有用的工具。在自噬过程中，吞噬体伸长形成双膜囊泡，但其闭合背后的机制尚不清楚。在这里，作者开发了一种自噬实验，并发现运输成分CHMP2A作为吞噬体关闭调节剂所需的内体分选复合物的作用。
The mechanism of phagophore closure remains unclear due to technical limitations in distinguishing unclosed and closed autophagosomal membranes. Here, we report the HaloTag-LC3 autophagosome completion assay that specifically detects phagophores, nascent autophagosomes, and mature autophagic structures. Using this assay, we identify the endosomal sorting complexes required for transport (ESCRT)-III component CHMP2A as a critical regulator of phagophore closure. During autophagy, CHMP2A translocates to the phagophore and regulates the separation of the inner and outer autophagosomal membranes to form double-membrane autophagosomes. Consistently, inhibition of the AAA-ATPase VPS4 activity impairs autophagosome completion. The ESCRT-mediated membrane abscission appears to be a critical step in forming functional autolysosomes by preventing mislocalization of lysosome-associated membrane glycoprotein 1 to the inner autophagosomal membrane. Collectively, our work reveals a function for the ESCRT machinery in the final step of autophagosome formation and provides a useful tool for quantitative analysis of autophagosome biogenesis and maturation. During autophagy, phagophores elongate to form double-membrane vesicles but the mechanism behind their closure is unknown. Here, the authors develop an autophagy assay and find a role for the endosomal sorting complexes required for transport component CHMP2A as a phagophore closure regulator.
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DOI：
10.1083/jcb.152.4.657
发表时间：
2001-02-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mizushima N;Yamamoto A;Hatano M;Kobayashi Y;Kabeya Y;Suzuki K;Tokuhisa T;Ohsumi Y;Yoshimori T
通讯作者：
Yoshimori T
影响因子：
16
作者：
Ganley IG;Wong PM;Gammoh N;Jiang X
通讯作者：
Jiang X
影响因子：
13.3
作者：
Knorr RL;Lipowsky R;Dimova R
通讯作者：
Dimova R
影响因子：
64.8
作者：
Olmos, Yolanda;Hodgson, Lorna;Mantell, Judith;Verkade, Paul;Carlton, Jeremy G.
通讯作者：
Carlton, Jeremy G.
影响因子：
4.5
作者：
Roudier, N;Lefebvre, C;Legouis, R
通讯作者：
Legouis, R