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The catalytic module that carries out ubiquitin (Ub) transfer remains unknown. Here we identify and characterize a distinct E3 module within human UBR4 consisting of a \u2018hemiRING\u2019 zinc finger, a helical-rich UBR zinc-finger interacting (UZI) subdomain, and an N-terminal region that can serve as an affinity factor for the E2 conjugating enzyme (E2). The structure of an E2\u2013E3 complex provides atomic-level insight into the specificity determinants of the hemiRING toward the cognate E2s UBE2A\/UBE2B. Via an allosteric mechanism, the UZI subdomain modestly activates the Ub-loaded E2 (E2\u223cUb). We propose attenuated activation is complemented by the intrinsically high lysine reactivity of UBE2A, and their cooperation imparts a reactivity profile important for substrate specificity and optimal degradation kinetics. These findings reveal the mechanistic underpinnings of a neuronal N-degron E3, its specific recruitment of UBE2A, and highlight the underappreciated architectural diversity of cross-brace domains with Ub E3 activity.<\/jats:p>","DOI":"10.1038\/s41594-023-01192-4","type":"journal-article","created":{"date-parts":[[2024,1,5]],"date-time":"2024-01-05T11:03:32Z","timestamp":1704452612000},"page":"351-363","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":36,"title":["UBE2A and UBE2B are recruited by an atypical E3 ligase module in UBR4"],"prefix":"10.1038","volume":"31","author":[{"given":"Lucy","family":"Barnsby-Greer","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7202-400X","authenticated-orcid":false,"given":"Peter D.","family":"Mabbitt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc-Andre","family":"Dery","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel R.","family":"Squair","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3347-6625","authenticated-orcid":false,"given":"Nicola T.","family":"Wood","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederic","family":"Lamoliatte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6011-8647","authenticated-orcid":false,"given":"Sven M.","family":"Lange","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7052-9802","authenticated-orcid":false,"given":"Satpal","family":"Virdee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,1,5]]},"reference":[{"key":"1192_CR1","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1073\/pnas.1816596116","volume":"116","author":"A Varshavsky","year":"2019","unstructured":"Varshavsky, A. 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