Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation
Whole genome screen reveals a novel relationship between Wolbachia levels and Drosophila host translation
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DOI：
10.1371/journal.ppat.1007445
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发表时间：
2018-11-01
期刊：
影响因子：
6.7
通讯作者：
Lehmann, Ruth
中科院分区：
文献类型：
--
作者：
Grobler, Yolande;Yun, Chi Y.;Lehmann, Ruth
沃尔巴克氏体是一种细胞内细菌，可感染多种昆虫宿主。蚊子等昆虫是登革热、西尼罗河和寨卡等许多毁灭性人类病毒的载体。值得注意的是，沃尔巴克氏体感染使昆虫宿主对许多虫媒病毒具有抵抗力，从而使昆虫失去作为媒介的作用。为了有效地利用沃尔巴克氏体作为对抗媒介传播病毒的工具，需要更好地了解宿主与沃尔巴克氏体的关系。为了研究沃尔巴克氏体与昆虫的相互作用，我们使用了沃尔巴克氏体/果蝇模型，该模型为研究宿主与病原体的相互作用提供了一个遗传上易于处理的系统。我们将全基因组 RNAi 筛选与新型高通量荧光原位杂交 (FISH) 测定相结合，以检测沃尔巴克氏体感染的果蝇细胞系 JW18 中沃尔巴克氏体水平的变化。当 RNAi 敲低时，1117 个基因改变了沃尔巴克氏体的水平，其中 329 个基因增加了沃尔巴克氏体的水平，788 个基因降低了沃尔巴克氏体的水平。使用体外 RNAi、果蝇突变体和 DNA qPCR 检测沃尔巴克氏体对深度二次筛选中的命中进行验证。确定了一组不同的宿主基因网络来调节沃尔巴克氏体水平，并意外地揭示了宿主翻译成分（例如核糖体和翻译起始因子）的扰动导致沃尔巴克氏体水平增加，无论是在体外使用RNAi还是在体内使用突变体和基于化学的翻译抑制测定。这项工作为沃尔巴克氏体-宿主翻译相互作用提供了证据，并加强了我们对沃尔巴克氏体-宿主细胞内关系的一般理解。
Wolbachia is an intracellular bacterium that infects a remarkable range of insect hosts. Insects such as mosquitos act as vectors for many devastating human viruses such as Dengue, West Nile, and Zika. Remarkably, Wolbachia infection provides insect hosts with resistance to many arboviruses thereby rendering the insects ineffective as vectors. To utilize Wolbachia effectively as a tool against vector-borne viruses a better understanding of the host-Wolbachia relationship is needed. To investigate Wolbachia-insect interactions we used the Wolbachia/Drosophila model that provides a genetically tractable system for studying host-pathogen interactions. We coupled genome-wide RNAi screening with a novel high-throughput fluorescence in situ hybridization (FISH) assay to detect changes in Wolbachia levels in a Wolbachia-infected Drosophila cell line JW18. 1117 genes altered Wolbachia levels when knocked down by RNAi of which 329 genes increased and 788 genes decreased the level of Wolbachia. Validation of hits included in depth secondary screening using in vitro RNAi, Drosophila mutants, and Wolbachia-detection by DNA qPCR. A diverse set of host gene networks was identified to regulate Wolbachia levels and unexpectedly revealed that perturbations of host translation components such as the ribosome and translation initiation factors results in increased Wolbachia levels both in vitro using RNAi and in vivo using mutants and a chemical-based translation inhibition assay. This work provides evidence for Wolbachia-host translation interaction and strengthens our general understanding of the Wolbachia-host intracellular relationship.