Cryo-electron Tomography Reveals the Roles of FliY in Helicobacter pylori Flagellar Motor Assembly.
Cryo-electron Tomography Reveals the Roles of FliY in Helicobacter pylori Flagellar Motor Assembly.
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冷冻电子断层扫描揭示 FliY 在幽门螺杆菌鞭毛运动组件中的作用
DOI：
10.1128/msphere.00944-21
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发表时间：
2022-02-23
期刊：
影响因子：
4.8
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Lu P;Zhang H;Gao Y;Jia X;Liu Z;Wang D;Au SWN;Zhang Q
幽门螺杆菌是与胃病相关的主要危险因素。鞭毛马达是幽门螺杆菌最重要的毒力因子之一。摘要幽门螺杆菌在胃部疾病中起致病作用。幽门螺杆菌的致病性取决于其在胃内的定植能力。FliY是一种独特的鞭毛电机开关成分，在一些细菌中与经典的FliG、FliM和FLIN开关蛋白共存，已被证明是鞭毛形成所必需的。然而，FliY在幽门螺杆菌鞭毛马达组装中的功能重要性还不是很清楚。在这里，我们应用冷冻电子断层扫描和亚断层图像平均分析了表达FliY的N-末端或C-末端结构域的野生型、fliY缺失突变体和互补突变体的鞭毛马达的原位结构。全长FliY或其C-末端结构域的丢失中断了完整C环和可溶性输出装置的形成，以及钩丝和鞭毛细丝的形成。与Fly Y C-末端结构域互补，恢复了鞭毛运动的所有缺失成分。综上所述，这些结果为了解FliY，特别是其C-末端结构域在幽门螺杆菌鞭毛运动组装中的作用提供了结构方面的见解。重要性幽门螺杆菌是与胃病相关的主要危险因素。鞭毛马达是幽门螺杆菌最重要的毒力因子之一。然而，幽门螺杆菌鞭毛马达的组装机制尚不完全清楚。以前的报告主要描述了鞭毛的总体结构，但没有侧重于其具体成分。在这里，我们重点研究幽门螺杆菌鞭毛C环蛋白FliY。我们通过冷冻电子断层扫描和亚断层图像平均，直接显示了幽门螺杆菌野生型和FliY N/C末端互补菌株的鞭毛结构。我们的结果表明，FliY或其C-末端结构域的缺失会导致C环的丢失，而FliY N-末端的缺失不会影响C-环的组装和鞭毛结构。我们的结果直接证明了C-环蛋白FliY，特别是其C-末端结构域，在幽门螺杆菌的运动组装和鞭毛形成过程中发挥着不可或缺的作用。本研究将加深我们对幽门螺杆菌致病机制的认识。
Helicobacter pylori is the major risk factor related with gastric diseases. Flagellar motor is one of the most important virulence factors in H. pylori. ABSTRACT Helicobacter pylori plays a causative role in gastric diseases. The pathogenicity of H. pylori depends on its ability to colonize the stomach guided by motility. FliY is a unique flagellar motor switch component coexisting with the classical FliG, FliM, and FliN switch proteins in some bacteria and has been shown to be essential for flagellation. However, the functional importance of FliY in H. pylori flagellar motor assembly is not well understood. Here, we applied cryo-electron tomography and subtomogram averaging to analyze the in situ structures of flagellar motors from wild-type strain, fliY-null mutant and complementation mutants expressing the N-terminal or C-terminal domain of FliY. Loss of full-length FliY or its C-terminal domain interrupted the formation of an intact C ring and soluble export apparatus, as well as the hook and flagellar filaments. Complementation with FliY C-terminal domain restored all these missing components of flagellar motor. Taken together, these results provide structural insights into the roles of FliY, especially its C-terminal domain in flagellar motor assembly in H. pylori. IMPORTANCE Helicobacter pylori is the major risk factor related with gastric diseases. Flagellar motor is one of the most important virulence factors in H. pylori. However, the assembly mechanism of H. pylori flagellar motor is not fully understood yet. Previous report mainly described the overall structures of flagellum but had not focused on its specific components. Here, we focus on H. pylori flagellar C-ring protein FliY. We directly visualize the flagellar structures of H. pylori wild-type and FliY N-/C-terminal complementary strains by cryo-electron tomography and subtomogram averaging. Our results show that deletion of FliY or its C-terminal domain causes the loss of C ring, whereas deletion of FliY N-terminal does not affect C-ring assembly and flagellar structures. Our results provide direct evidence that C-ring protein FliY, especially its C-terminal domain, plays an indispensable role in H. pylori motor assembly and flagellar formation. This study will deepen our understanding about H. pylori pathogenesis.
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影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1073/pnas.1110111108
发表时间：
2011-10-11
影响因子：
11.1
作者：
Paul, Koushik;Brunstetter, Duncan;Blair, David F.
通讯作者：
Blair, David F.
影响因子：
5.2
作者：
Lam, Wendy Wai-Ling;Sun, Kailei;Au, Shannon Wing-Ngor
通讯作者：
Au, Shannon Wing-Ngor
影响因子：
3
作者：
Kremer, JR;Mastronarde, DN;McIntosh, JR
通讯作者：
McIntosh, JR
影响因子：
3.2
作者：
Lowder, BJ;Duyvesteyn, MD;Blair, DF
通讯作者：
Blair, DF