High-through put isolation of functional mutants of actin using Dictyostelium expression system

使用盘基网柄菌表达系统高通量分离肌动蛋白功能突变体

• 批准号: 22570163
• 负责人: WAKABAYASHI Takeyuki
• 金额: $ 2.91万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22570163/
• 关键词: アクチン; ミオシン; 細胞分化; 収縮タンパク質; 機能的変異株; 細胞性粘菌; X 線結晶解析; ATPase; アクチン重合; アクチン・ミオシン相互作用; X線結晶解析; アクチン変異体; ディクチオ型粘菌; 胞子形成能; アクチンフィラメント; アクチン重合能

Actin filaments are found in all eukaryotic cells and are essentialfor many of their movements including cell migration and cell division. We found that the cell behavior of Dictyostelium cells changed, when the vector for the expression of mutant actin was incorporated. Using this phenomenon, functional mutants of actin were isolated efficiently. It was found that the tyrosine-143 of actin is important for actin assembly and the activation of myosin ATPase. The crystal structures of such mutants were determined.

肌动蛋白丝存在于所有真核细胞中，对于细胞迁移和细胞分裂等许多运动至关重要。我们发现，当掺入表达突变肌动蛋白的载体时，盘基网柄菌细胞的细胞行为发生了变化。利用这种现象，有效分离了肌动蛋白的功能突变体。研究发现肌动蛋白的酪氨酸143对于肌动蛋白的组装和肌球蛋白ATP酶的激活很重要。确定了此类突变体的晶体结构。

项目成果

Electron microscopic evidence for the myosin head lever arm mechanism inhydrated myosin filaments using the gas environmental chambe

使用气体环境室的肌球蛋白头杠杆臂机制水合肌球蛋白丝的电子显微镜证据

• 发表时间: 2011
• 期刊: Biochem. Biophys. Res. Commun
• 作者: H. Minoda;T. Okabe;Y. Inayoshi;T. Minakata;Y. Miyauchi;M. Tanokura;E. Katayama;T. Wakabayashi;T. Akimoto;*H. Sugi
• 通讯作者: *H. Sugi

Changes of biochemical properties of Dictyostelium actin induced by mutation of tyrosine-143

酪氨酸143突变引起盘基网柄菌肌动蛋白生化特性的变化

• 发表时间: 2012
• 作者: 五味渕由貴; 上田太郎; 若林健之
• 通讯作者: 若林健之

Dominant negative mutant actins identified in flightless Drosophila can be classified into three classes, including perturbation of the tropomyosin-troponin system

在不会飞的果蝇中发现的显性负突变肌动蛋白可分为三类，包括原肌球蛋白-肌钙蛋白系统的扰动

• DOI: 10.1074/jbc.m109.059881
• 发表时间: 2010
• 期刊: J. Biol. Chem.
• 作者: Noguchi;T. Q. P.;Y. Gomibuchi;K. Murakami;H. Ueno;K. Hirose;T. Wakabayashi;and T. Q. P. Uyeda
• 通讯作者: and T. Q. P. Uyeda

Structural Basis of Actin Polymerization, ATPase activation and Delayed Phosphate Release

肌动蛋白聚合、ATP 酶激活和延迟磷酸盐释放的结构基础

• 发表时间: 2010
• 作者: K. Murakami;T. Yasunaga;T. Q. P. Noguchi;Y. Gomibuchi;K. Ngo;T. Q. P. Uyeda;and T. Wakabayashi
• 通讯作者: and T. Wakabayashi

How actin assembly triggers activation of ATP hydrolysis

肌动蛋白组装如何触发 ATP 水解的激活

• 发表时间: 2010
• 作者: K. Murakami;T. Yasunaga;T. Q. P. Noguchi;T. Q. P. Uyeda & T . Wakabayashi
• 通讯作者: T. Q. P. Uyeda & T . Wakabayashi