New clearance system of abnormal protein in Polyglutammine disease

多聚谷氨酰胺病异常蛋白的新清除系统

• 批准号: 13480258
• 负责人: NAKAYAMA Keiichi
• 金额: $ 9.66万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480258/
• 关键词: MJD; UFD2; VPC; polyglutamine Disease; Ubiquitylation; MJD1; ノックアウトマウス

Machado-Joseph disease (MJD)/Spinocerebellar ataxia type 3 (SCA3) is neurodegenerative disease which is caused by polyglutamine expansion in a responsible gene product, MJD1/Ataxin3. MJD1 has now been shown to undergo ubiquitylation and degradation by proteasome-dependent pathway. MJD1 with expanded polygiutamine tract was more resistant to degradation than normal MJD1. We established an in vitro system of ubiquitylation of MJD1, thereby biochemically purified activity to mediate polyubiquitylation of MJD1 from rabbit reticulocyte lysate. An AAA-family ATPase VCP was isolated from the active fraction, and found to binds to MJD1. Furthermore, UFD2a, a mammalian ubiquitin-chain assembly factor (E4), associated with VCP and induced polyubiquitylation of MJD1. UFD2a markedly promoted ubiquitylation and degradation of MJD1 with expanded polyglutamine tract, resulting in the clearance of MJD1 protein. In contrast, dominant-negative mutant UFD2a reduced the degradation rate of MJD1, leading to the formation of intracellular aggregation. In Drosophila model, overexpression of UFD2a significantly suppressed the neurodegeneration induced by expression of MJD1 with expanded polyglutamine tract. These findings suggest that E4 is a ratelimiting factor of degradation of pathologic polyglutamine-containing proteins, and may give a potential tool for gene therapy to control the clinical conditions of MJD

Machado-Joseph病(MJD)/脊髓小脑性共济失调3型（SCA3）是一种神经退行性疾病，由负责基因产物MJD1/Ataxin3中的聚谷氨酰胺扩增引起。MJD1现已被证明通过蛋白酶体依赖途径进行泛素化和降解。与正常MJD1相比，具有扩张多聚氰胺束的MJD1具有更强的抗降解能力。我们建立了MJD1的体外泛素化体系，从而从兔网织细胞裂解液中生化纯化了介导MJD1泛素化的活性。从活性组分中分离出一个aaa家族atp酶VCP，发现它与MJD1结合。此外，哺乳动物泛素链组装因子（E4） UFD2a与VCP相关，并诱导MJD1的多泛素化。UFD2a通过扩大聚谷氨酰胺通道，显著促进MJD1的泛素化和降解，导致MJD1蛋白被清除。相比之下，显性阴性突变体UFD2a降低了MJD1的降解速率，导致细胞内聚集的形成。在果蝇模型中，过表达UFD2a可显著抑制MJD1表达引起的多聚谷氨酰胺束扩张神经退行性变。这些研究结果表明，E4是病理性含多聚谷氨酰胺蛋白降解的限速因子，可能为基因治疗控制MJD的临床状况提供潜在的工具

项目成果

Kanematsu, T. et al.: "Role of the PLC-related, catalytically inactive protein p130 in GABA_A receptor function"EMBO J.. 21. 1004-1011 (2002)

Kanematsu, T. 等人：“PLC 相关的催化失活蛋白 p130 在 GABA_A 受体功能中的作用”EMBO J.. 21. 1004-1011 (2002)

Shimoda K. et. al.: "Tyk2 is required for the induction and nuclear translocation of Daxx which regulates IFN-α-induced suppression of B lymphocyte formation"J. Immunol.. 169. 4707-4711 (2002)

Shimoda K. 等人：“Tyk2 是 Daxx 的诱导和核易位所必需的，它调节 IFN-α 诱导的 B 淋巴细胞形成抑制”J.Immunol.. 169. 4707-4711 (2002)

Shimoda K. et. al.: "Partial impairment of interleukin-12 (IL-12) and 1L-18 signaling in Tyk2-deficient mice"Blood. 99. 2094-2099 (2002)

下田K.等。

Nagahama, H.: "Spatial and temporal expression patterns of the cyclin-dependent kinase (CDK) inhibitors p27^<Kip1> and p57^<Kip2> during mouse development"Anat. Embryol. 203. 77-87 (2001)

Nagahama, H.：“小鼠发育过程中细胞周期蛋白依赖性激酶 (CDK) 抑制剂 p27^<Kip1> 和 p57^<Kip2> 的空间和时间表达模式”Anat。

Masuda T. et al.: "Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma : modulation of malignant phenotype by Skp2 overexpression, possibly via p27"Cancer Res.. 62. 3819-3825 (2002)

Masuda T. 等人：“胃癌中 S 期激酶相关蛋白 2 (Skp2) 基因表达的临床和生物学意义：Skp2 过表达调节恶性表型，可能通过 p27”Cancer Res.. 62. 3819-