Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Nat Struct Mol Biol. 2020 Jun;27(6):589-597. doi: 10.1038/s41594-020-0429-1. Epub 2020 May 18.
The Vibrio parahaemolyticus T3SS effector VopQ targets host-cell V-ATPase, resulting in blockage of autophagic flux and neutralization of acidic compartments. Here, we report the cryo-EM structure of VopQ bound to the V subcomplex of the V-ATPase. VopQ inserts into membranes and forms an unconventional pore while binding directly to subunit c of the V-ATPase membrane-embedded subcomplex V. We show that VopQ arrests yeast growth in vivo by targeting the immature V subcomplex in the endoplasmic reticulum (ER), thus providing insight into the observation that VopQ kills cells in the absence of a functional V-ATPase. VopQ is a bacterial effector that has been discovered to inhibit a host-membrane megadalton complex by coincidentally binding its target, inserting into a membrane and disrupting membrane potential. Collectively, our results reveal a mechanism by which bacterial effectors modulate host cell biology and provide an invaluable tool for future studies on V-ATPase-mediated membrane fusion and autophagy.
副溶血性弧菌 T3SS 效应物 VopQ 靶向宿主细胞 V-ATPase，导致自噬流阻断和酸性隔室中和。在这里，我们报告了与 V-ATPase V 亚基复合物结合的 VopQ 的冷冻电镜结构。VopQ 插入膜中并形成非常规孔，同时直接与 V-ATPase 膜嵌入亚基 V 的亚基 c 结合。我们表明，VopQ 通过靶向内质网 (ER) 中的未成熟 V 亚基在体内阻止酵母生长，从而为观察到的在缺乏功能 V-ATPase 的情况下 VopQ 杀死细胞提供了线索。VopQ 是一种细菌效应物，已被发现通过巧合地结合其靶标、插入膜中和破坏膜电位来抑制宿主膜兆道尔顿复合物。总的来说，我们的结果揭示了细菌效应物调节宿主细胞生物学的机制，并为未来关于 V-ATPase 介导的膜融合和自噬的研究提供了宝贵的工具。
