title: Biochemical and Functional Characterization of RNF213 (Mysterin) R4810K, a Susceptibility Mutation of Moyamoya Disease, in Angiogenesis In Vitro and In Vivo.
abstract: BACKGROUND
  P.R4810K of RNF213 (mysterin: rs112735431), which is an AAA(+) ATPase, is the susceptibility polymorphism for moyamoya disease (MMD) in East Asians. However, the role of RNF213 R4810K in the etiology of MMD is unknown.
  
  
  RESULTS
  To clarify the role of RNF213 in known angiogenic pathways, RNF213 expression was analyzed in endothelial cells (ECs) treated with several angiogenic and antiangiogenic factors, including interferons (IFNs). RNF213 was upregulated by IFN-β through signal transducer and activator of transcription x in the promoter and mediated antiangiogenic activity of IFN-β. RNF213 wild-type (WT) overexpression could not lower angiogenesis without IFN-β, but RNF213 R4810K overexpression could. To correlate biochemical function as ATPase and the role of RNF213 oligomer formation with antiangiogenic activity, we investigated the effects of mutations in the AAA(+) module. A mutation of the Walker B motif (WEQ), which stabilizes oligomerization, inhibited angiogenesis, but AAA(+) module deletion, which cannot initiate oligomerization, did not. Intriguingly, R4810K, similar to WEQ, decreased ATPase activity, suggesting its antiangiogenic activity through stabilizing oligomers. To confirm the antiangiogenic effect of RNF213 upregulation in vivo, vascular EC- or smooth muscle cell-specific Rnf213 R4757K (R4810K ortholog) or WT transgenic (Tg) mice were exposed to hypoxia. Cerebral angiogenesis by hypoxia was suppressed in EC-specific Rnf213 R4757K Tg mice, whereas it was not suppressed in other mice.
  
  
  CONCLUSIONS
  This study suggests the importance of inflammatory signals as environmental factors and R4810K carriers for susceptibility to cerebral hypoxia. A specific inhibitor of ATP binding to the first AAA(+) could be a promising therapeutic candidate for MMD.
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.).
authors.forename: Hatasu
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Department of Neurosurgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan (Y.M., Y.T., S.M.).
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Matsuda
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.) Department of Preventive Medicine, St Marianna University School of Medicine, Kawasaki, Japan (T.H.).
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Hitomi
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.).
authors.forename: Hiroko
authors.initials: H
authors.lastname: Okuda
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.).
authors.forename: Hirotomo
authors.initials: H
authors.lastname: Shioi
authors.affiliation: Department of Systems Science, Graduate School of Informatics, Kyoto University, Kyoto, Japan (T.M., H.I.).
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Matsuda
authors.affiliation: Department of Systems Science, Graduate School of Informatics, Kyoto University, Kyoto, Japan (T.M., H.I.).
authors.forename: Hirohiko
authors.initials: H
authors.lastname: Imai
authors.affiliation: Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, Kyoto, Japan (M.S., D.T.).
authors.forename: Masakatsu
authors.initials: M
authors.lastname: Sone
authors.affiliation: Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, Kyoto, Japan (M.S., D.T.).
authors.forename: Daisuke
authors.initials: D
authors.lastname: Taura
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.).
authors.forename: Kouji H
authors.initials: KH
authors.lastname: Harada
authors.affiliation: Radiation Biology Center, Kyoto University, Kyoto, Japan (T.H.).
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Habu
authors.affiliation: Department of Neurosurgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan (Y.M., Y.T., S.M.).
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Takagi
authors.affiliation: Department of Neurosurgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan (Y.M., Y.T., S.M.).
authors.forename: Susumu
authors.initials: S
authors.lastname: Miyamoto
authors.affiliation: Department of Health and Environmental Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan (H.K., T.H., H.O., H.S., K.H.H., A.K.).
authors.forename: Akio
authors.initials: A
authors.lastname: Koizumi
mesh_terms: D000251:Adenosine Triphosphatases; D000255:Adenosine Triphosphate / Q000378:metabolism; D020816:Amino Acid Motifs; D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002465:Cell Movement; D049109:Cell Proliferation; D002536:Cerebral Arteries / Q000187:drug effects / Q000201:enzymology*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D020022:Genetic Predisposition to Disease; D057809:HEK293 Cells; D006367:HeLa Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D000860:Hypoxia / Q000150:complications; D018836:Inflammation Mediators / Q000494:pharmacology; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009072:Moyamoya Disease / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D009154:Mutation; D032389:Myocytes, Smooth Muscle / Q000201:enzymology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010641:Phenotype; D013997:Time Factors; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D043925:Angiogenesis Inducing Agents; D020533:Angiogenesis Inhibitors; D018836:Inflammation Mediators; D000255:Adenosine Triphosphate; C557998:RNF213 protein, human; C583291:RNF213 protein, mouse; D044767:Ubiquitin-Protein Ligases; D000251:Adenosine Triphosphatases
keywords: ATPase; RNF213; interferon; moyamoya disease; transgenic mouse