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Title: The moyamoya disease susceptibility variant RNF213 R4810K (rs112735431) induces genomic instability by mitotic abnormality

Highlights: •Overexpression of RNF213 R4810K inhibited cell proliferation. •Overexpression of RNF213 R4810K had the time of mitosis 4-fold and mitotic failure. •R4810K formed a complex with MAD2 more readily than wild-type. •iPSECs from the MMD patients had elevated mitotic failure compared from the control. •RNF213 R4810K induced mitotic abnormality and increased risk of aneuploidy. -- 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, 13.1more » ± 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.« less
Authors:
 [1] ;  [2] ; ; ;  [1] ;  [3] ; ;  [4] ; ; ; ; ; ;  [3] ; ;  [5] ;  [6] ;  [5] ;  [4] ;  [1]
  1. Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto (Japan)
  2. Radiation Biology Center, Kyoto University, Kyoto (Japan)
  3. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto (Japan)
  4. Department of Medicine and Clinical Science, Graduate School of Medicine, Kyoto University, Kyoto (Japan)
  5. Department of Neurosurgery, Graduate School of Medicine, Kyoto University, Kyoto (Japan)
  6. Faculty of Life Sciences, Kyoto Sangyo University, Kyoto (Japan)
Publication Date:
OSTI Identifier:
22242115
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 439; Journal Issue: 4; Other Information: Copyright (c) 2013 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ABLATION; ANEUPLOIDY; CELL CYCLE; CELL PROLIFERATION; COMPARATIVE EVALUATIONS; COMPLEXES; DISEASES; FIBROBLASTS; HAZARDS; HELA CELLS; INHIBITION; INSTABILITY; MITOSIS; MORPHOLOGY; PATIENTS; STEM CELLS