Shi Xingqi, Halder Partho, Yavuz Halenur, Jahn Reinhard, Shuman Howard A
Department of Microbiology, University of Chicago, Chicago, IL 60637;
Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, 37077 Goettingen, Germany.
Proc Natl Acad Sci U S A. 2016 Aug 2;113(31):8807-12. doi: 10.1073/pnas.1608755113. Epub 2016 Jul 19.
Legionella pneumophila, the Gram-negative pathogen causing Legionnaires' disease, infects host cells by hijacking endocytic pathways and forming a Legionella-containing vacuole (LCV) in which the bacteria replicate. To promote LCV expansion and prevent lysosomal targeting, effector proteins are translocated into the host cell where they alter membrane traffic. Here we show that three of these effectors [LegC2 (Legionella eukaryotic-like gene C2)/YlfB (yeast lethal factor B), LegC3, and LegC7/YlfA] functionally mimic glutamine (Q)-SNARE proteins. In infected cells, the three proteins selectively form complexes with the endosomal arginine (R)-SNARE vesicle-associated membrane protein 4 (VAMP4). When reconstituted in proteoliposomes, these proteins avidly fuse with liposomes containing VAMP4, resulting in a stable complex with properties resembling canonical SNARE complexes. Intriguingly, however, the LegC/SNARE hybrid complex cannot be disassembled by N-ethylmaleimide-sensitive factor. We conclude that LegCs use SNARE mimicry to divert VAMP4-containing vesicles for fusion with the LCV, thus promoting its expansion. In addition, the LegC/VAMP4 complex avoids the host's disassembly machinery, thus effectively trapping VAMP4 in an inactive state.
嗜肺军团菌是引起军团病的革兰氏阴性病原体,它通过劫持内吞途径感染宿主细胞,并形成一个含军团菌的液泡(LCV),细菌在其中进行复制。为促进LCV的扩张并防止其被溶酶体靶向,效应蛋白会转运到宿主细胞中,在那里它们会改变膜运输。在这里,我们表明其中三种效应蛋白[LegC2(军团菌类真核基因C2)/YlfB(酵母致死因子B)、LegC3和LegC7/YlfA]在功能上模拟谷氨酰胺(Q)-SNARE蛋白。在受感染的细胞中,这三种蛋白与内体精氨酸(R)-SNARE囊泡相关膜蛋白4(VAMP4)选择性地形成复合物。当在蛋白脂质体中重构时,这些蛋白会与含有VAMP4的脂质体 avidly融合,形成一种性质类似于典型SNARE复合物的稳定复合物。然而,有趣的是,LegC/SNARE杂交复合物不能被N-乙基马来酰亚胺敏感因子拆解。我们得出结论,LegC蛋白利用SNARE模拟来转移含VAMP4的囊泡与LCV融合,从而促进其扩张。此外,LegC/VAMP4复合物避开了宿主的拆解机制,从而有效地将VAMP4捕获在无活性状态。