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Characterization of disease-associated mutations has been complicated by the enormous size of RNF213. Here, we present the cryo-EM structure of mouse RNF213. The structure reveals the intricate fold of the 584 kDa protein, comprising an N-terminal stalk, a dynein-like core with six ATPase units, and a multidomain E3 module. Collaboration with UbcH7, a cysteine-reactive E2, points to an unexplored ubiquitin-transfer mechanism that proceeds in a RING-independent manner. Moreover, we show that pathologic MMD mutations cluster in the composite E3 domain, likely interfering with substrate ubiquitination. In conclusion, the structure of RNF213 uncovers a distinct type of an E3 enzyme, highlighting the growing mechanistic diversity in ubiquitination cascades. Our results also provide the molecular framework for investigating the emerging role of RNF213 in lipid metabolism, hypoxia, and angiogenesis.<\/jats:p>","DOI":"10.7554\/elife.56185","type":"journal-article","created":{"date-parts":[[2020,6,18]],"date-time":"2020-06-18T07:00:17Z","timestamp":1592463617000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.56185","source":"Crossref","is-referenced-by-count":135,"title":["Moyamoya disease factor RNF213 is a giant E3 ligase with a dynein-like core and a distinct ubiquitin-transfer mechanism"],"prefix":"10.7554","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0293-1063","authenticated-orcid":true,"given":"Juraj","family":"Ahel","sequence":"first","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anita","family":"Lehner","sequence":"additional","affiliation":[{"name":"Vienna BioCenter Core Facilities, Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonia","family":"Vogel","sequence":"additional","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Schleiffer","sequence":"additional","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anton","family":"Meinhart","sequence":"additional","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5276-5633","authenticated-orcid":true,"given":"David","family":"Haselbach","sequence":"additional","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1582-6924","authenticated-orcid":true,"given":"Tim","family":"Clausen","sequence":"additional","affiliation":[{"name":"Research Institute of Molecular Pathology (IMP), Vienna BioCenter, Vienna, Austria"},{"name":"Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4374","published-online":{"date-parts":[[2020,6,23]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1107\/S2059798318006551","article-title":"Real-space refinement in PHENIX for cryo-EM and crystallography","volume":"74","author":"Afonine","year":"2018","journal-title":"Acta Crystallographica Section D Structural Biology"},{"key":"bib2","article-title":"CrossLinkingVisualizer","author":"Ahel","year":"2020","unstructured":"Ahel J. 2020. 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