| Literature DB >> 30030437 |
Yoshinori Takahashi1, Haiyan He2, Zhenyuan Tang2, Tatsuya Hattori2, Ying Liu2, Megan M Young2, Jacob M Serfass3, Longgui Chen2, Melat Gebru2, Chong Chen2, Carson A Wills2, Jennifer M Atkinson2, Han Chen4, Thomas Abraham4,5, Hong-Gang Wang6,7.
Abstract
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.Entities:
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Year: 2018 PMID： 30030437 PMCID： PMC6054611 DOI： 10.1038/s41467-018-05254-w
Source DB: PubMed Journal: Nat Commun ISSN： 2041-1723 Impact factor: 14.919