Degradation process of expanded polyglutamine protein in SCA6
| Project/Area Number |
16590814
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | Tokyo Medical And Dental University |
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Principal Investigator |
FUJIGASAKI Hiroto Tokyo Medical And Dental University, Department of Neurology, part time lecturer, 大学院・医歯学総合研究科, 非常勤講師 (40345286)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | SCA6 / polyglutamine / ubiquitin / splisosome |
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| Research Abstract |
Background : Spinocerebellar ataxia type 6 (SCA6) is one of the autosomal dominant cerebellar ataxias caused by an abnormally expanded CAG repeat in the alpha1A-calcium channel (CACNA1A) gene. To investigate the pathophysiology of the disease, we established a cellular model for SCA6 using Tet-off system.
Method : Expression vectors (pTRE2-hyg) encoding C-terminal fragments of CACNA1A containing a normal (13Q) or an expanded (28Q) polyglutamine stretch were transfected to Tet-off PC12 cells and Hyg resistant clones were selected under the manufacture's protocol. Expression levels of truncated proteins with 13Q or 28Q were confirmed by Western blot analyses. Subcellular localization of these proteins was evaluated by immunohistochemistry.
Results : Inducible stable cell lines expressing truncated CACNA1A proteins were established. Expression levels were regulated by doxycyclin. Immunohistochemical analyses revealed that 28Q cells formed cytoplasmic aggregates including a mutant protein as we demonstrated in SCA6 brains previously.
Conclusion : We established a cellular model for SCA6 using Tet-off system. 28Q cells form aggregates like Purkinje cells in SCA6 brains. Our SCA6 model is useful to observe the forming process of cytoplasmic aggregates induced by CACNA1A containing an expanded polyglutamine.
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Report
(3 results)
Research Products
(9 results)
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[Journal Article] An autosomal dominant cerebellar ataxia linked to chromosome 16q22.1 is associated with a single-nucleotide substitution in the 5' untranslated region of the gene encoding a protein with spectrin repeat and Rho guanine-nucleotide exchange factor domains.2005
Author(s)
Ishikawa K, Toru S, Tsunemi T, Li M, Kobayashi K, Yokota T, Amino T, Owada K, Fuiigasaki H, Sakamoto M, Tomimitsu H, Takashima M, Kumagai J, Noguchi Y, Kawashima Y, Ohkoshi N, Ishida G, Gomyoda M, Yoshida M Hashizume Y, Saito Y, Murayama S, Yamanouchi H, Mizutani T, Kondo I, Toda T, Mizusawa, H.
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Journal Title
Am J Hum Genet 77
Pages: 280-296
Description
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