Laura and Isaac Perlmutter Cancer Center, New York University Langone Medical Center, New York University, New York, NY, USA.
Department of Medical Biophysics, University of Toronto, Toronto, Canada.
Life Sci Alliance. 2022 Feb 8;5(5). doi: 10.26508/lsa.202000807. Print 2022 May.
Single-nucleotide polymorphisms (SNPs) in , 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 , 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, , is a dominant-negative mutant that decreases ubiquitylation globally. By contrast, MMD-associated SNPs do not affect ATPase activity. Our results suggest that decreased RNF213 E3 ligase activity is central to MMD pathogenesis.
单核苷酸多态性（SNPs）位于 ，编码一个具有 AAA+ATPase 和 RING E3 结构域的 591kD 蛋白，与一种罕见的常染色体显性脑血管疾病，烟雾病（MMD）有关。与 MMD 相关的 SNPs 主要定位于 ，一些影响 RING 结构域中的保守残基。虽然 MMD 的常染色体显性遗传最容易通过 RNF213 获得功能来解释，但 RNF213 催化的泛素化类型以及 MMD 相关 SNPs 对其 E3 连接酶活性的影响仍不清楚。我们发现 RNF213 使用 E2 连接酶 UBE2D2 和 UBE2L3 来催化不同的泛素化事件。RNF213-UBE2D2 在体外催化 K6 和在较小程度上 K48 依赖性多泛素化，而 RNF213-UBE2L3 催化 K6-、K11-和 K48 依赖性多泛素化事件。与 MMD 相关的 SNPs 编码具有降低的 E3 活性的蛋白质，最常见的 MMD 等位基因 ，是一种降低泛素化全局水平的显性负突变体。相比之下，与 MMD 相关的 SNPs 不影响 ATPase 活性。我们的结果表明，RNF213 E3 连接酶活性的降低是 MMD 发病机制的核心。
