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In situ structure of the Caulobacter crescentus flagellar motor and visualization of binding of a CheY-homolog.
Molecular Microbiology ( IF 2.6 ) Pub Date : 2020-05-25 , DOI: 10.1111/mmi.14525
Florian M Rossmann 1 , Isabelle Hug 2 , Matteo Sangermani 2 , Urs Jenal 2 , Morgan Beeby 1
Affiliation  

Bacterial flagellar motility is controlled by the binding of CheY proteins to the cytoplasmic switch complex of the flagellar motor, resulting in changes in swimming speed or direction. Despite its importance for motor function, structural information about the interaction between effector proteins and the motor are scarce. To address this gap in knowledge, we used electron cryotomography and subtomogram averaging to visualize such interactions inside Caulobacter crescentus cells. In C. crescentus, several CheY homologs regulate motor function for different aspects of the bacterial lifestyle. We used subtomogram averaging to image binding of the CheY family protein CleD to the cytoplasmic Cring switch complex, the control center of the flagellar motor. This unambiguously confirmed the orientation of the motor switch protein FliM and the binding of a member of the CheY protein family to the outside rim of the C ring. We also uncovered previously unknown structural elaborations of the alphaproteobacterial flagellar motor, including two novel periplasmic ring structures, and the stator ring harboring eleven stator units, adding to our growing catalog of bacterial flagellar diversity.

中文翻译:

新月柄杆菌鞭毛运动的原位结构和 CheY 同源物结合的可视化。

细菌鞭毛运动是通过 CheY 蛋白与鞭毛运动细胞质开关复合物的结合来控制的,从而导致游动速度或方向的变化。尽管它对运动功能很重要,但有关效应蛋白和运动之间相互作用的结构信息却很少。为了解决这一知识空白,我们使用电子冷冻断层扫描和亚断层扫描平均来可视化新月柄杆菌细胞内的这种相互作用。在C. crescentus中,几种 CheY 同源物调节细菌生活方式不同方面的运动功能。我们使用亚断层图平均对 CheY 家族蛋白 CleD 与细胞质 Cring 开关复合体(鞭毛运动的控制中心)的结合进行成像。这明确证实了运动开关蛋白 FliM 的方向以及 CheY 蛋白家族成员与 C 环外缘的结合。我们还发现了以前未知的α变形菌鞭毛运动的结构细节,包括两个新颖的周质环结构,以及包含十一个定子单元的定子环,增加了我们不断增长的细菌鞭毛多样性目录。
更新日期:2020-05-25
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