title: Lipidation of the LC3/GABARAP family of autophagy proteins relies on a membrane-curvature-sensing domain in Atg3.
abstract: The components supporting autophagosome growth on the cup-like isolation membrane are likely to be different from those found on closed and maturing autophagosomes. The highly curved rim of the cup may serve as a functionally required surface for transiently associated components of the early acting autophagic machinery. Here we demonstrate that the E2-like enzyme, Atg3, facilitates LC3/GABARAP lipidation only on membranes exhibiting local lipid-packing defects. This activity requires an amino-terminal amphipathic helix similar to motifs found on proteins targeting highly curved intracellular membranes. By tuning the hydrophobicity of this motif, we can promote or inhibit lipidation in vitro and in rescue experiments in Atg3-knockout cells, implying a physiologic role for this stress detection. The need for extensive lipid-packing defects suggests that Atg3 is designed to work at highly curved membranes, perhaps including the limiting edge of the growing phagophore.
authors.affiliation: 1] Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA [2].
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Nath
authors.affiliation: 1] Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA [2].
authors.forename: Julia
authors.initials: J
authors.lastname: Dancourt
authors.affiliation: 1] Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA [2].
authors.forename: Vladimir
authors.initials: V
authors.lastname: Shteyn
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Puente
authors.affiliation: Departments of Neurology, Pathology and Cell Biology, Columbia University, New York 10032, USA.
authors.forename: Wendy M
authors.initials: WM
authors.lastname: Fong
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
authors.forename: Shanta
authors.initials: S
authors.lastname: Nag
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
authors.forename: Joerg
authors.initials: J
authors.lastname: Bewersdorf
authors.affiliation: Departments of Neurology, Pathology and Cell Biology, Columbia University, New York 10032, USA.
authors.forename: Ai
authors.initials: A
authors.lastname: Yamamoto
authors.affiliation: Institut de Pharmacologie Moléculaire et Cellulaire, CNRS et université de Nice, 06103, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Antonny
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Melia
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D000071193:Autophagy-Related Protein 7; D000071190:Autophagy-Related Protein 8 Family; D000071183:Autophagy-Related Proteins; D002462:Cell Membrane / Q000201:enzymology*; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008081:Liposomes; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010714:Phosphatidylethanolamines / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D013312:Stress, Physiological; D014162:Transfection; D044764:Ubiquitin-Activating Enzymes / Q000235:genetics / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D000071193:Autophagy-Related Protein 7; D000071190:Autophagy-Related Protein 8 Family; D000071183:Autophagy-Related Proteins; D003598:Cytoskeletal Proteins; D008081:Liposomes; D008565:Membrane Proteins; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D010714:Phosphatidylethanolamines; D044764:Ubiquitin-Activating Enzymes; D044763:Ubiquitin-Conjugating Enzymes; C480861:GABARAP protein, mouse; C430503:GABARAPL1 protein, human; C498698:GABARAPL2 protein, human; C526109:Anxa3 protein, rat; C480860:Map1lc3b protein, mouse; C483858:phosphatidylethanolamine; C509349:ATG7 protein, human; C568853:Atg3 protein, mouse
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