{"doi":"10.1101/2021.05.14.444175","title":"Structural Basis for Membrane Recruitment of ATG16L1 by WIPI2 in Autophagy","abstract":"ABSTRACT 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 bladed β-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 β-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12–5-16L1 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 ATG12–5-16L1 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.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2021,"id":220783,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9526,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":273460,"name":"Chunmei Chang","orcid":"0000-0002-5607-7985","position":1,"is_corresponding":false},{"id":488527,"name":"C. Alexander Boecker","orcid":"0000-0001-9701-5273","position":2,"is_corresponding":false},{"id":237128,"name":"Thomas G. Flower","orcid":"0000-0002-7890-6473","position":3,"is_corresponding":false},{"id":237129,"name":"Cosmo Z. Buffalo","orcid":"0000-0002-8187-4660","position":4,"is_corresponding":false},{"id":237132,"name":"Xuefeng Ren","orcid":"0000-0002-4822-4316","position":5,"is_corresponding":false},{"id":820471,"name":"Andrea KH Stavoe","orcid":"0000-0002-4073-4565","position":6,"is_corresponding":false},{"id":258944,"name":"Erika L.F. Holzbaur","orcid":"0000-0001-5389-4114","position":7,"is_corresponding":false},{"id":237133,"name":"James H. Hurley","orcid":"0000-0001-5054-5445","position":8,"is_corresponding":false},{"id":582772,"name":"Lisa Strong","orcid":"0000-0002-4293-8131","position":0,"is_corresponding":true}],"reference_count":50,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:53:50.838581Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}