TY - JOUR
T1 - VPS37A directs ESCRT recruitment for phagophore closure
AU - Takahashi, Yoshinori
AU - Liang, Xinwen
AU - Hattori, Tatsuya
AU - Tang, Zhenyuan
AU - He, Haiyan
AU - Chen, Han
AU - Liu, Xiaoming
AU - Abraham, Thomas
AU - Imamura-Kawasawa, Yuka
AU - Buchkovich, Nicholas J.
AU - Young, Megan M.
AU - Wang, Hong Gang
PY - 2019/10/7
Y1 - 2019/10/7
N2 - The process of phagophore closure requires the endosomal sorting complex required for transport III (ESCRT-III) subunit CHMP2A and the AAA ATPase VPS4, but their regulatory mechanisms remain unknown. Here, we establish a FACS-based HaloTag-LC3 autophagosome completion assay to screen a genome-wide CRISPR library and identify the ESCRT-I subunit VPS37A as a critical component for phagophore closure. VPS37A localizes on the phagophore through the N-terminal putative ubiquitin E2 variant domain, which is found to be required for autophagosome completion but dispensable for ESCRT-I complex formation and the degradation of epidermal growth factor receptor in the multivesicular body pathway. Notably, loss of VPS37A abrogates the phagophore recruitment of the ESCRT-I subunit VPS28 and CHMP2A, whereas inhibition of membrane closure by CHMP2A depletion or VPS4 inhibition accumulates VPS37A on the phagophore. These observations suggest that VPS37A coordinates the recruitment of a unique set of ESCRT machinery components for phagophore closure in mammalian cells.
AB - The process of phagophore closure requires the endosomal sorting complex required for transport III (ESCRT-III) subunit CHMP2A and the AAA ATPase VPS4, but their regulatory mechanisms remain unknown. Here, we establish a FACS-based HaloTag-LC3 autophagosome completion assay to screen a genome-wide CRISPR library and identify the ESCRT-I subunit VPS37A as a critical component for phagophore closure. VPS37A localizes on the phagophore through the N-terminal putative ubiquitin E2 variant domain, which is found to be required for autophagosome completion but dispensable for ESCRT-I complex formation and the degradation of epidermal growth factor receptor in the multivesicular body pathway. Notably, loss of VPS37A abrogates the phagophore recruitment of the ESCRT-I subunit VPS28 and CHMP2A, whereas inhibition of membrane closure by CHMP2A depletion or VPS4 inhibition accumulates VPS37A on the phagophore. These observations suggest that VPS37A coordinates the recruitment of a unique set of ESCRT machinery components for phagophore closure in mammalian cells.
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U2 - 10.1083/jcb.201902170
DO - 10.1083/jcb.201902170
M3 - Article
C2 - 31519728
SN - 0021-9525
VL - 218
SP - 3336
EP - 3354
JO - The Journal of cell biology
JF - The Journal of cell biology
IS - 10
ER -