title: The moyamoya disease susceptibility variant RNF213 R4810K (rs112735431) induces genomic instability by mitotic abnormality.
abstract: Moyamoya disease (MMD) is a cerebrovascular disease characterized by occlusive lesions in the Circle of Willis. The RNF213 R4810K polymorphism increases susceptibility to MMD. In the present study, we characterized phenotypes caused by overexpression of RNF213 wild type and R4810K variant in the cell cycle to investigate the mechanism of proliferation inhibition. Overexpression of RNF213 R4810K in HeLa cells inhibited cell proliferation and extended the time of mitosis 4-fold. Ablation of spindle checkpoint by depletion of mitotic arrest deficiency 2 (MAD2) did not shorten the time of mitosis. Mitotic morphology in HeLa cells revealed that MAD2 colocalized with RNF213 R4810K. Immunoprecipitation revealed an RNF213/MAD2 complex: R4810K formed a complex with MAD2 more readily than RNF213 wild-type. Desynchronized localization of MAD2 was observed more frequently during mitosis in fibroblasts from patients (n=3, 61.0 ± 8.2%) compared with wild-type subjects (n=6, 13.1 ± 7.7%; p<0.01). Aneuploidy was observed more frequently in fibroblasts (p<0.01) and induced pluripotent stem cells (iPSCs) (p<0.03) from patients than from wild-type subjects. Vascular endothelial cells differentiated from iPSCs (iPSECs) of patients and an unaffected carrier had a longer time from prometaphase to metaphase than those from controls (p<0.05). iPSECs from the patients and unaffected carrier had significantly increased mitotic failure rates compared with controls (p<0.05). Thus, RNF213 R4810K induced mitotic abnormalities and increased risk of genomic instability.
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Hitomi
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Habu
authors.forename: Hatasu
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Hiroko
authors.initials: H
authors.lastname: Okuda
authors.forename: Kouji H
authors.initials: KH
authors.lastname: Harada
authors.forename: Kenji
authors.initials: K
authors.lastname: Osafune
authors.forename: Daisuke
authors.initials: D
authors.lastname: Taura
authors.forename: Masakatsu
authors.initials: M
authors.lastname: Sone
authors.forename: Isao
authors.initials: I
authors.lastname: Asaka
authors.forename: Tomonaga
authors.initials: T
authors.lastname: Ameku
authors.forename: Akira
authors.initials: A
authors.lastname: Watanabe
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kasahara
authors.forename: Tomomi
authors.initials: T
authors.lastname: Sudo
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Shiota
authors.forename: Hirokuni
authors.initials: H
authors.lastname: Hashikata
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Takagi
authors.forename: Daisuke
authors.initials: D
authors.lastname: Morito
authors.forename: Susumu
authors.initials: S
authors.lastname: Miyamoto
authors.forename: Kazuwa
authors.initials: K
authors.lastname: Nakao
authors.forename: Akio
authors.initials: A
authors.lastname: Koizumi
mesh_terms: D000251:Adenosine Triphosphatases; D020022:Genetic Predisposition to Disease; D014644:Genetic Variation; D042822:Genomic Instability; D005838:Genotype; D006367:HeLa Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D064246:Mad2 Proteins / Q000235:genetics / Q000378:metabolism; D008938:Mitosis / Q000235:genetics*; D009072:Moyamoya Disease / Q000235:genetics*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
chemical_list: C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; C557998:RNF213 protein, human; D044767:Ubiquitin-Protein Ligases; D000251:Adenosine Triphosphatases
keywords: Genomic instability; MAD2; Mitotic phase; Moyamoya disease; Rs112735431; iPS cells