Nucleocytoplasmic shuttling of cytoskeletal proteins : molecular mechanism and biological significance

细胞骨架蛋白的核质穿梭：分子机制和生物学意义

• 批准号: 22870017
• 负责人: KUMETA Masahiro
• 金额: $ 2.01万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22870017/
• 关键词: 細胞・組織; 分子細胞生物学; 核膜孔複合体; 細胞骨格; 核輸送; アクチニン; スペクトリン

In this study we focused on the communication between cytoskeleton and the nucleus, which is achieved by the dynamic shuttling of cytoskeletal components into the nucleus. Nuclear pore complex(NPC) serves as a hydrophobic selective gate of nucleocytoplasmic molecular transport, enabling small molecules(~ 40kDa) and nuclear transport receptors(e. g., importins and exportins) to pass through the nuclear membrane. However, the detailed selectivity property of NPC still remained unknown. First, we demonstrated the nucleocytoplasmic shuttling of several cytoskeletal proteins such as spectrin and actinin by immunostaining of cells treated with Leptomycin B, a selective inhibitor for CRM1-dependent nuclear export. Co-precipitation and quantitative PCR assays revealed their role in fundamental nuclear functions such as gene expression. In vitro nuclear transport assay using digitonin-treated cytoplasm-depleted cells clearly showed their interesting property to spontaneously migrate into the nucleus. Because these proteins possess a characteristic amphiphilic motif in their 3D structure, it was speculated that this amphiphilic motif plays an important role in the molecular passage through the hydrophobic barrier of the pore. Surface hydrophobicity measurement by using

在这项研究中，我们专注于细胞骨架和细胞核之间的沟通，这是通过动态穿梭的细胞骨架成分进入细胞核。核孔复合物（NPC）是核质分子转运的疏水选择性门，使小分子（~ 40 kDa）和核转运受体（如：例如，在一个实施例中，输入和输出）通过核膜。然而，NPC的详细选择性性质仍然未知。首先，我们证明了几种细胞骨架蛋白，如血影蛋白和辅肌动蛋白的核质穿梭的Leptomycin B，CRM 1依赖性核输出的选择性抑制剂处理的细胞的免疫染色。共沉淀和定量PCR分析揭示了它们在基因表达等基本核功能中的作用。使用毛地黄皂苷处理的细胞质耗尽细胞的体外核转运试验清楚地显示了它们自发迁移到细胞核中的有趣特性。由于这些蛋白质在其3D结构中具有特征性的两亲性基序，因此推测该两亲性基序在通过孔的疏水屏障的分子通道中起重要作用。表面疏水性测量

项目成果

生体の科学-「核小体の単離・精製とその構造」

生物科学-《核仁及其结构的分离纯化》

• 发表时间: 2011
• 作者: Emilie Louvet;粂田昌宏;竹安邦夫
• 通讯作者: 竹安邦夫

Karyopherin-independent nuclear transport of large cytoskeletal proteins

大细胞骨架蛋白的不依赖核转运蛋白的核转运

• 发表时间: 2011
• 作者: Masahiro Kumeta;Shige H. Yoshimura and Kunio Takeyasu
• 通讯作者: Shige H. Yoshimura and Kunio Takeyasu

核膜孔のサイズバリアを越えて-巨大タンパク質の核内移行メカニズム

跨越核孔径屏障——大蛋白质的核输入机制

• 发表时间: 2011
• 作者: 粂田昌宏;山口秀輝;吉村成弘;竹安邦夫
• 通讯作者: 竹安邦夫

Dynamic regulation of nuclear functions by subcellular scaffolding proteins

亚细胞支架蛋白对核功能的动态调节

• 发表时间: 2010
• 作者: Uchihashi;T.;Kodera;N.;Ando;T.;Masahiro Kumeta
• 通讯作者: Masahiro Kumeta

Spectrin-repeat proteins in the nucleus : molecular mechanism and biological significance of the nuclear localization

细胞核中的血影蛋白重复蛋白：核定位的分子机制和生物学意义

• 发表时间: 2010
• 作者: Masahiro Kumeta;Shige H. Yoshimura and Kunio Takeyasu
• 通讯作者: Shige H. Yoshimura and Kunio Takeyasu