An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure

被引：255

作者：

Takahashi, Yoshinori ^{[1
]}

He, Haiyan ^{[1
]}

Tang, Zhenyuan ^{[1
]}

Hattori, Tatsuya ^{[1
]}

Liu, Ying ^{[1
]}

Young, Megan M. ^{[1
]}

Serfass, Jacob M. ^{[2
]}

Chen, Longgui ^{[1
]}

Gebru, Melat ^{[1
]}

Chen, Chong ^{[1
]}

Wills, Carson A. ^{[1
]}

Atkinson, Jennifer M. ^{[1
]}

Chen, Han ^{[3
]}

Abraham, Thomas ^{[3
,4
]}

Wang, Hong-Gang ^{[1
,2
]}

机构：

[1] Penn State Coll Med, Dept Pediat, Hershey, PA 17033 USA

[2] Penn State Coll Med, Dept Pharmacol, Hershey, PA 17033 USA

[3] Penn State Coll Med, Microscopy Imaging Facil, Hershey, PA 17033 USA

[4] Penn State Coll Med, Dept Neural & Behav Sci, Hershey, PA 17033 USA

来源：

NATURE COMMUNICATIONS | 2018年 / 9卷

关键词：

CASPASE-8; ACTIVATION; CELL-SURVIVAL; PROTEIN; MECHANISMS; FISSION; LC3; TRAFFICKING; DISSECTION; MATURATION; INDUCTION;

D O I：

10.1038/s41467-018-05254-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：13

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