Search for inhibitory factor for polyglutamine protein ubioquitylation

寻找多聚谷氨酰胺蛋白泛素化的抑制因子

• 批准号: 18590929
• 负责人: IWATA Atsushi
• 金额: $ 2.34万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590929/
• 关键词: polyglutamine; ubiquitin; proteasome; 神経変性疾患

This year, we tested if any of the ubiquitin like proteins could modify polyglutamine proteins. We cloned FAT10, ubl5, FubI, Isg15 and urm1 from cDNA library and made overexpression constructs for mammalian expression. We co-expressed these constructs with polyglutamine proteins and pulled down pQ with immunoprecipitation method. Immunodetection of attached proteins revealed that FAT10 was covalently attached to pulled-down pQ proteins. This was further confirmed by pulling down pQ under denatured condition to detect FAT10 covalently bound to pQs. FAT10 is known to be homologous to ubiquitin abd known to covalently conjugated to lysine residues. We tested various amounts of FAT10 with ubiquitin co-expressed to find FAT10 affects ubiquitylation of PQ protein. By overexpressing FAT10, degradation of pQ proteins are accelerated. These data suggested that FAT10 might play and important role in pQ pathogenesis. We are now confirming the data and preparing for publication.

今年，我们测试了是否有任何泛素样蛋白可以修饰聚谷氨酰胺蛋白。我们从cDNA文库中克隆了FAT10、ubl5、FubI、Isg15和urm1，并构建了用于哺乳动物表达的过表达载体。我们将这些构建体与多聚谷氨酰胺蛋白共表达，并通过免疫沉淀方法拉低pQ。免疫检测显示，FAT10共价连接到下拉的pQ蛋白。这通过在变性条件下拉下pQ以检测与pQ共价结合的FAT10来进一步证实。已知FAT10与泛素同源，并已知与赖氨酸残基共价缀合。我们测试了不同数量的FAT10与泛素共表达，以发现FAT10影响PQ蛋白的泛素化。通过过表达FAT10，加速了pQ蛋白的降解。提示FAT10可能在pQ发病中起重要作用。我们现在正在确认数据并准备发表。

项目成果

神経変性疾患とオートファジー

神经退行性疾病和自噬

• DOI: 10.32206/jkpum.131.02.115
• 发表时间: 2022
• 期刊: Journal of Kyoto Prefectural University of Medicine
• 作者: Fan Q.;Takarada-Iemata M.;Hori O;服部 剛志;渡邊義久
• 通讯作者: 渡邊義久

Assembly of Lysine 63-linked Ubiquitin Conjugates by Phosphorylated {alpha}-Synuclein Implies Lewy Body Biogenesis.

通过磷酸化 {α}-突触核蛋白组装赖氨酸 63 连接的泛素缀合物意味着路易体生物发生。

• 发表时间: 2007
• 期刊: Journal of Biological Chemistry 282
• 作者: Liu C;Fei E;Jia N;Wang H;Tao R;Iwata A;Nukina N;Zhou J;Wang G.
• 通讯作者: Wang G.

HDAC6 and autophagy

HDAC6 和自噬

• 发表时间: 2007
• 作者: Liu C;Fei E;Jia N;Wang H;Tao R;Iwata A;Nukina N;Zhou J;Wang G.;Watanabe S;Nakamura M;Nakashima I;岩田淳;Nakashima I;岩田淳;Nakamura M;岩田淳;Watanabe S;岩田淳;Misu T;Takahashi T;Atsushi Iwata
• 通讯作者: Atsushi Iwata

オートファジーと神経疾患

自噬与神经系统疾病

• 发表时间: 2006
• 期刊: 医学のあゆみ 216
• 作者: Liu C;Fei E;Jia N;Wang H;Tao R;Iwata A;Nukina N;Zhou J;Wang G.;Watanabe S;Nakamura M;Nakashima I;岩田淳
• 通讯作者: 岩田淳

HDAC6とオートファジー

HDAC6 和自噬

• 发表时间: 2007
• 作者: Liu C;Fei E;Jia N;Wang H;Tao R;Iwata A;Nukina N;Zhou J;Wang G.;Watanabe S;Nakamura M;Nakashima I;岩田淳;Nakashima I;岩田淳;Nakamura M;岩田淳;Watanabe S;岩田淳
• 通讯作者: 岩田淳