Energy and information transduction of bacteria flagellar motor of chimeric cell

嵌合细胞细菌鞭毛运动的能量和信息转导

• 批准号: 20370056
• 负责人: ISHIJIMA Akihiko
• 金额: $ 13.31万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20370056/
• 关键词: バクテリア; 回転計測; モータータンパク; 温度; トルク; イメージング; GFP; FRAP; エバネッセント

An Escherichia coli cell transduces extracellular stimuli sensed by chemoreceptors to the state of an intracellular signal molecule, which regulates switching of the rotational direction of the flagellar motors from counterclockwise (CCW) to clockwise (CW). Here, we performed high-speed imaging of flagellar motor rotation and showed that the switching of two different motors on a cell is controlled coordinately by an intracellular signal protein, phosphorylated CheY (CheY-P). The switching is highly coordinated, with a sub-second delay between motors that is correlated with the distance of each motor from the chemoreceptor patch, which is localized at the cell pole. The coordinated switching was disordered by the expression of constitutively active CheY protein, which mimics the CW-rotation stimulating function. Also, the coordinated switching was required the CheZ, which is a phosphatase for CheY-P. Our results suggest that a transient increase and decrease in the concentration of a signal protein from the chemoreceptor patch, which is probably a wave-like change in under a second, triggers and regulates the coordinated switching of flagellar motors. We firstly measured the propagation of intracellular signaling in a single bacterial cell by applying the simultaneous measurement for the rotation of multiple flagellar motors.

大肠杆菌细胞将化学感受器感知的细胞外刺激转变成细胞内信号分子的状态，从而调节鞭毛马达的旋转方向从逆时针 (CCW) 到顺时针 (CW) 的切换。在这里，我们对鞭毛运动旋转进行了高速成像，结果表明细胞上两种不同运动的切换是由细胞内信号蛋白磷酸化 CheY (CheY-P) 协调控制的。这种切换是高度协调的，电机之间有亚秒级的延迟，该延迟与每个电机与化学感受器贴片（位于细胞极处）的距离相关。协调性转换因模拟 CW 旋转刺激功能的组成型活性 CheY 蛋白的表达而紊乱。此外，协调转换需要 CheZ，它是 CheY-P 的磷酸酶。我们的结果表明，化学感受器斑块信号蛋白浓度的短暂增加和减少（可能是一秒钟内的波状变化）触发并调节鞭毛马达的协调切换。我们首先通过同时测量多个鞭毛马达的旋转来测量单个细菌细胞中细胞内信号的传播。

项目成果

光学顕微鏡を用いた細菌べん毛モーター1分子の観察および計測

使用光学显微镜观察和测量单个细菌鞭毛运动分子

• 发表时间: 2009
• 作者: Hajiome Fukuoka;Tomoyuki Wada;Seiji Kojima; Akihiko Ishijima;Michio Homma;福岡創,石島秋彦
• 通讯作者: 福岡創,石島秋彦

Turnover of subunit-exchange in functioning bacterial flagellar motor

功能性细菌鞭毛运动中亚基交换的周转

• 发表时间: 2009
• 作者: Hajime Fukuoka;Shun Terasawa;Hiroto Takahashi;Yuichi Inoue;Akihiko Ishijima
• 通讯作者: Akihiko Ishijima

高速カメラおよび微小ビーズを用いた大腸菌べん毛モーターの回転計測:走化性シグナルに対するべん毛モーター回転運動の解析

使用高速相机和微珠测量大肠杆菌鞭毛电机的旋转：分析响应趋化信号的鞭毛电机旋转

• 发表时间: 2010
• 作者: 井上裕一;石島秋彦
• 通讯作者: 石島秋彦

FRAP analysis for visualization of exchange of rotor component in functional bacteria flagellar motor

功能性细菌鞭毛运动中转子组件交换可视化的 FRAP 分析

• 发表时间: 2008
• 作者: Hajime Fukuoka;Shun Terasawa;Yuichi Inoue;Takeharu Nagai and Akihiko Ishijima
• 通讯作者: Takeharu Nagai and Akihiko Ishijima

TIRFM observation of chemoreceptor cluster and chemo sensory complex in E. coli cell membrane

大肠杆菌细胞膜化学感受器簇和化学感觉复合物的 TIRFM 观察

• 发表时间: 2008
• 作者: Takehiko Inaba;Satomi Banno;Hajime Fukuoka;Akihiko Ishijima and Ikuro Kawaaishi
• 通讯作者: Akihiko Ishijima and Ikuro Kawaaishi