Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy
Autophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double-membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12–5–16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethano...
Main Authors: | , , , , , , , , , |
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Language: | English |
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eLife Sciences Publications Ltd
2021-09-01
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Online Access: | https://elifesciences.org/articles/70372 |
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author | Lisa M Strong Chunmei Chang Julia F Riley C Alexander Boecker Thomas G Flower Cosmo Z Buffalo Xuefeng Ren Andrea KH Stavoe Erika LF Holzbaur James H Hurley |
author_facet | Lisa M Strong Chunmei Chang Julia F Riley C Alexander Boecker Thomas G Flower Cosmo Z Buffalo Xuefeng Ren Andrea KH Stavoe Erika LF Holzbaur James H Hurley |
author_sort | Lisa M Strong |
collection | DOAJ |
description | Autophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double-membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12–5–16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethanolamine in the growing autophagosomal membrane, known as the phagophore. ATG12–5–16L1 is recruited to the phagophore by a subset of the phosphatidylinositol 3-phosphate-binding seven-bladedß -propeller WIPI proteins. We determined the crystal structure of WIPI2d in complex with the WIPI2 interacting region (W2IR) of ATG16L1 comprising residues 207–230 at 1.85 Å resolution. The structure shows that the ATG16L1 W2IR adopts an alpha helical conformation and binds in an electropositive and hydrophobic groove between WIPI2 ß-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12–5–16 L1 and the conjugation of the ATG8 protein LC3B to synthetic membranes. Interface mutants show a decrease in starvation-induced autophagy. Comparisons across the four human WIPIs suggest that WIPI1 and 2 belong to a W2IR-binding subclass responsible for localizing ATG12–5–16 L1 and driving ATG8 lipidation, whilst WIPI3 and 4 belong to a second W34IR-binding subclass responsible for localizing ATG2, and so directing lipid supply to the nascent phagophore. The structure provides a framework for understanding the regulatory node connecting two central events in autophagy initiation, the action of the autophagic PI 3-kinase complex on the one hand and ATG8 lipidation on the other. |
first_indexed | 2024-04-11T10:48:36Z |
format | Article |
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institution | Directory Open Access Journal |
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language | English |
last_indexed | 2024-04-11T10:48:36Z |
publishDate | 2021-09-01 |
publisher | eLife Sciences Publications Ltd |
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series | eLife |
spelling | doaj.art-3680dcace3464a88937af5f1d613cc8d2022-12-22T04:28:58ZengeLife Sciences Publications LtdeLife2050-084X2021-09-011010.7554/eLife.70372Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagyLisa M Strong0https://orcid.org/0000-0002-4293-8131Chunmei Chang1https://orcid.org/0000-0002-5607-7985Julia F Riley2https://orcid.org/0000-0001-8518-9786C Alexander Boecker3https://orcid.org/0000-0001-9701-5273Thomas G Flower4https://orcid.org/0000-0002-7890-6473Cosmo Z Buffalo5Xuefeng Ren6Andrea KH Stavoe7https://orcid.org/0000-0002-4073-4565Erika LF Holzbaur8https://orcid.org/0000-0001-5389-4114James H Hurley9https://orcid.org/0000-0001-5054-5445Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United StatesAligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United States; Department of Physiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, United StatesAligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United States; Department of Physiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United StatesDepartment of Neurobiology and Anatomy, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, United StatesAligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United States; Department of Physiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, United StatesAutophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double-membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12–5–16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethanolamine in the growing autophagosomal membrane, known as the phagophore. ATG12–5–16L1 is recruited to the phagophore by a subset of the phosphatidylinositol 3-phosphate-binding seven-bladedß -propeller WIPI proteins. We determined the crystal structure of WIPI2d in complex with the WIPI2 interacting region (W2IR) of ATG16L1 comprising residues 207–230 at 1.85 Å resolution. The structure shows that the ATG16L1 W2IR adopts an alpha helical conformation and binds in an electropositive and hydrophobic groove between WIPI2 ß-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12–5–16 L1 and the conjugation of the ATG8 protein LC3B to synthetic membranes. Interface mutants show a decrease in starvation-induced autophagy. Comparisons across the four human WIPIs suggest that WIPI1 and 2 belong to a W2IR-binding subclass responsible for localizing ATG12–5–16 L1 and driving ATG8 lipidation, whilst WIPI3 and 4 belong to a second W34IR-binding subclass responsible for localizing ATG2, and so directing lipid supply to the nascent phagophore. The structure provides a framework for understanding the regulatory node connecting two central events in autophagy initiation, the action of the autophagic PI 3-kinase complex on the one hand and ATG8 lipidation on the other.https://elifesciences.org/articles/70372autophagymitophagyparkinson's diseasex-ray crystallographyvesicle reconstitutionLC3 |
spellingShingle | Lisa M Strong Chunmei Chang Julia F Riley C Alexander Boecker Thomas G Flower Cosmo Z Buffalo Xuefeng Ren Andrea KH Stavoe Erika LF Holzbaur James H Hurley Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy eLife autophagy mitophagy parkinson's disease x-ray crystallography vesicle reconstitution LC3 |
title | Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy |
title_full | Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy |
title_fullStr | Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy |
title_full_unstemmed | Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy |
title_short | Structural basis for membrane recruitment of ATG16L1 by WIPI2 in autophagy |
title_sort | structural basis for membrane recruitment of atg16l1 by wipi2 in autophagy |
topic | autophagy mitophagy parkinson's disease x-ray crystallography vesicle reconstitution LC3 |
url | https://elifesciences.org/articles/70372 |
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