TY - JOUR T1 - MMD-associated <em>RNF213</em> SNPs encode dominant-negative alleles that globally impair ubiquitylation JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.202000807 VL - 5 IS - 5 SP - e202000807 AU - Abhishek Bhardwaj AU - Robert S Banh AU - Wei Zhang AU - Sachdev S Sidhu AU - Benjamin G Neel Y1 - 2022/05/01 UR - https://www.life-science-alliance.org/content/5/5/e202000807.abstract N2 - Single-nucleotide polymorphisms (SNPs) in RNF213, which encodes a 591-kD protein with AAA+ ATPase and RING E3 domains, are associated with a rare, autosomal dominant cerebrovascular disorder, moyamoya disease (MMD). MMD-associated SNPs primarily localize to the C-terminal region of RNF213, and some affect conserved residues in the RING domain. Although the autosomal dominant inheritance of MMD could most easily explained by RNF213 gain-of-function, the type of ubiquitylation catalyzed by RNF213 and the effects of MMD-associated SNPs on its E3 ligase activity have remained unclear. We found that RNF213 uses the E2-conjugating enzymes UBE2D2 and UBE2L3 to catalyze distinct ubiquitylation events. RNF213-UBED2 catalyzes K6 and, to a lesser extent, K48-dependent poly-ubiquitylation in vitro, whereas RNF213-UBE2L3 catalyzes K6-, K11-, and K48-dependent poly-ubiquitylation events. MMD-associated SNPs encode proteins with decreased E3 activity, and the most frequent MMD allele, RNF213R4810K, is a dominant-negative mutant that decreases ubiquitylation globally. By contrast, MMD-associated RNF213 SNPs do not affect ATPase activity. Our results suggest that decreased RNF213 E3 ligase activity is central to MMD pathogenesis. ER -