Host Factors Required for Dengue and Yellow Fever Virus Amplification

描述（由申请人提供）： 登革热和黄热病病毒是引起出血热和死亡的重要人类病原体。目前，还没有针对这些病毒的批准的抗病毒药物。由于耐药病毒的快速出现，RNA病毒的抗病毒药物的开发已被证明是特别困难的。这主要是由于聚合酶的易错性，这导致病毒在选择压力下快速进化。针对宿主因子， 病毒没有任何遗传控制，应该减少或防止抗性突变体的发展，因为病毒没有宿主因子就不能发挥作用，或者因为需要更多的突变来赋予抗性。为了实现这一目标，我们已经开发了一种稳健的方法来鉴定在细胞培养中病毒扩增期间与病毒RNA结合的宿主因子。我们将使用这种方法来鉴定与登革热病毒和黄热病毒RNA结合的宿主因子。我们将确定哪些宿主因子将是开发抗病毒靶点的良好靶点。此外，我们还将确定宿主因子参与病毒生命周期的步骤，以及病毒感染期间该宿主因子的蛋白质-蛋白质和蛋白质-病毒RNA相互作用。这些研究将解决宿主因子在病毒扩增中的机制，并能够开发宿主因子抑制剂的高通量检测方法。 公共卫生相关性： RNA病毒迅速进化出对靶向病毒蛋白的抗病毒药物具有抗性的病毒。靶向宿主蛋白质的抗病毒药物将显著降低或防止耐药性。因此，我们将确定与登革热和黄热病病毒RNA相互作用的宿主因子，并确定它们在病毒生命周期中的作用，以确定有效的抗病毒靶点。

DESCRIPTION (provided by applicant): Dengue and yellow fever virus are significant human pathogens that cause hemorrhagic fever and fatalities. Currently, there are no approved antivirals for these viruses. The development of antivirals for RNA viruses has proven to be particularly difficult due to the rapid emergence of drug resistant viruses. This is primarily due to the error prone nature of the polymerase, which results in rapid evolution of the virus under selective pressure. Targeting a host factor for which the virus does not have any genetic control over should decrease or prevent development of resistant mutants either because the virus cannot function without the host factor or because more mutations are required to confer resistance. Towards this goal we have developed a robust method to identify host factors that are bound to the viral RNA during viral amplification i cell culture. We will use this method to identify host factors that are bound to the dengue virus and yellow fever virus RNA. We will identify which host factors will be good targets to develop antiviral targets. In addition, we will also determine the step in the viral life cycle the host fators participate in and the protein-protein and protein-viral RNA interactions for that host factor during viral infection. These studies will address the mechanism of the host factor in viral amplification and enable development of a high-throughput assay fro inhibitors of the host factor. PUBLIC HEALTH RELEVANCE: RNA viruses rapidly evolve resistant viruses to antiviral that target the viral proteins. An antivial that targets a host protein would significantly reduce or prevent drug resistance. Therefore, we will identify host factors that interact with dengue and yellow fever viral RNAs and determine their role in the viral life cycle in order to identify an effective antiviral target.