MMD-associated RNF213 SNPs encode dominant-negative alleles that globally impair ubiquitylation.

Abstract

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, RNF213 R4810K , 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.

• Authors: Abhishek Bhardwaj, Robert S Banh, Wei Zhang, Sachdev S Sidhu, Benjamin G Neel
• Journal: Life science alliance (Life Sci Alliance) Vol. 5 Issue 5 (05 2022) ISSN: 2575-1077 [Electronic] United States
• PMID: 35135845 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2022 Bhardwaj et al.

Chemical References

• Transcription Factors
• UBE2D2 protein, human
• UBE2L3 protein, human
• Ubiquitin-Conjugating Enzymes
• RNF213 protein, human
• Ubiquitin-Protein Ligases
• Adenosine Triphosphatases

Topics

• Adenosine Triphosphatases (genetics, metabolism)
• Alleles
• Genetic Predisposition to Disease
• HEK293 Cells
• HeLa Cells
• Humans
• Moyamoya Disease (genetics, pathology)
• Mutation
• Polymorphism, Single Nucleotide (genetics)
• Protein Domains (genetics)
• Transcription Factors (metabolism)
• Ubiquitin-Conjugating Enzymes (metabolism)
• Ubiquitin-Protein Ligases (genetics, metabolism)
• Ubiquitination (genetics)