Identification of New Protein Interactions between Dengue Fever Virus and Its Hosts, Human and Mosquito

The four divergent serotypes of dengue virus are the causative agents of dengue fever, dengue hemorrhagic fever and dengue shock syndrome. About two-fifths of the world's population live in areas where dengue is prevalent, and thousands of deaths are caused by the viruses every year. Dengue virus is transmitted from one person to another primarily by the yellow fever mosquito, Aedes aegypti. Recent studies have begun to define how the dengue viral proteins interact with host proteins to mediate viral replication and pathogenesis. A combined analysis of these studies, however, suggests that many virus-host protein interactions remain to be identified, especially for the mosquito host. In this study, we used high-throughput yeast two-hybrid screening to identify mosquito and human proteins that physically interact with dengue proteins. We tested each identified host protein against the proteins from all four serotypes of dengue to identify interactions that are conserved across serotypes. We further confirmed many of the interactions using co-affinity purification assays. As in other large-scale screens, we identified some previously detected interactions and many new ones, moving us closer to a complete host – dengue protein interactome. To help summarize and prioritize the data for further study, we combined our interactions with other published data and identified a subset of the host-dengue interactions that are now supported by multiple forms of evidence. These data should be useful for understanding the interplay between dengue and its hosts and may provide candidates for drug targets and vector control strategies.登革病毒（Dengue virus）的四种不同血清型，是登革热、登革出血热及登革休克综合征的致病病原体。全球约五分之二的人口居住在登革热流行区域，每年该病毒都会造成数千人死亡。登革病毒主要通过黄热病蚊媒——埃及伊蚊（Aedes aegypti）——在人际间传播。近期已有研究开始阐明登革病毒蛋白与宿主蛋白的相互作用机制，以此介导病毒复制与致病过程。然而，对这些研究的综合分析显示，仍有大量病毒-宿主蛋白相互作用有待发掘，针对蚊媒宿主的此类相互作用尤其如此。本研究采用高通量酵母双杂交筛选（high-throughput yeast two-hybrid screening）技术，鉴定出可与登革病毒蛋白发生物理相互作用的蚊媒及人类宿主蛋白。我们针对所有四种登革病毒血清型的蛋白，对每一个鉴定得到的宿主蛋白进行了测试，以筛选出在不同血清型间保守存在的相互作用。我们进一步通过共亲和纯化实验（co-affinity purification assays）验证了其中多数相互作用。与其他大规模筛选研究类似，本研究既鉴定出了此前已被报道的相互作用，也发现了大量全新的相互作用，使我们向完整绘制宿主-登革病毒蛋白相互作用组（protein interactome）的目标更近了一步。为辅助梳理数据并筛选出可供后续研究的重点方向，我们将本研究得到的相互作用数据与其他已发表的研究数据进行整合，鉴定出了目前有多类实验证据支持的宿主-登革病毒相互作用子集。本研究所得数据将有助于阐明登革病毒与其宿主之间的相互作用关系，亦可为药物靶点开发及媒介控制策略提供候选研究方向。