The activity of the conserved Atg12-Atg5-Atg16 complex is essential for autophagosome formation. However, little is known about its mechanism of action during this process. In our study we employed in vitro systems consisting of purified proteins and giant unilamellar vesicles (GUVs) or small liposomes to investigate membrane binding by the Atg12-Atg5-Atg16 complex and its interplay with the Atg8 conjugation system. We showed that Atg5 directly binds membranes and that this membrane binding is negatively regulated by Atg12 conjugation but activated by Atg16. Membrane binding by the Atg12-Atg5-Atg16 complex is required for efficient promotion of Atg8 lipidation. Additionally, we found that the Atg12-Atg5-Atg16 complex tethered vesicles in an Atg8-independent manner. In yeast, membrane binding by Atg5 is not required for its recruitment to the phagophore assembly site (PAS) but is essential for efficient promotion of autophagy and the cytoplasm-to-vacuole targeting (Cvt) pathway at a stage preceding Atg8 lipidation and autophagosome closure. Our findings provide new insights into the role of the Atg12-Atg5-Atg16 complex during autophagosome formation.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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