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

Yoshinori Takahashi; Haiyan He; Zhenyuan Tang; Tatsuya Hattori; Ying Liu; Megan M. Young; Jacob M. Serfass; Longgui Chen; Melat Gebru; Chong Chen; Carson A. Wills; Jennifer M. Atkinson; Han Chen; Thomas Abraham; Hong Gang Wang

doi:10.1038/s41467-018-05254-w

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

Yoshinori Takahashi, Haiyan He, Zhenyuan Tang, Tatsuya Hattori, Ying Liu, Megan M. Young, Jacob M. Serfass, Longgui Chen, Melat Gebru, Chong Chen, Carson A. Wills, Jennifer M. Atkinson, Han Chen, Thomas Abraham, Hong Gang Wang

Research output: Contribution to journal › Article › peer-review

66 Scopus citations

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.

Original language	English (US)
Article number	2855
Journal	Nature communications
Volume	9
Issue number	1
DOIs	https://doi.org/10.1038/s41467-018-05254-w
State	Published - Dec 1 2018

All Science Journal Classification (ASJC) codes

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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Takahashi, Yoshinori ; He, Haiyan ; Tang, Zhenyuan ; Hattori, Tatsuya ; Liu, Ying ; Young, Megan M. ; Serfass, Jacob M. ; Chen, Longgui ; Gebru, Melat ; Chen, Chong ; Wills, Carson A. ; Atkinson, Jennifer M. ; Chen, Han ; Abraham, Thomas ; Wang, Hong Gang. / An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure. In: Nature communications. 2018 ; Vol. 9, No. 1.

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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.",

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An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure. / Takahashi, Yoshinori; He, Haiyan; Tang, Zhenyuan; Hattori, Tatsuya; Liu, Ying; Young, Megan M.; Serfass, Jacob M.; Chen, Longgui; Gebru, Melat; Chen, Chong; Wills, Carson A.; Atkinson, Jennifer M.; Chen, Han; Abraham, Thomas ; Wang, Hong Gang.

In: Nature communications, Vol. 9, No. 1, 2855, 01.12.2018.

Research output: Contribution to journal › Article › peer-review

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AU - Abraham, Thomas

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