Nucleocapsid-Specific Small Molecule Inhibitors of the Bunyaviridae

布尼亚病毒科病毒包括一组A类和C类病毒种。布尼亚病毒科 这些病毒包括汉他病毒属的成员，以及裂谷热病毒和克里米亚-刚果病毒。 出血热病毒目前针对大多数这些病毒的治疗方法要么不存在，要么只是 边际效率。因此，鉴定和开发针对以下的另外的药物将是非常有益的： 复制所需的特定病毒靶点。本修订提案的目标是完善和使用一种测定方法， 用于鉴定破坏主要核衣壳蛋白功能的分子的系统 这些病毒，并进行筛选这些候选药物。核衣壳蛋白（N） 在病毒装配过程中的RNA降解过程中，以及在基因组复制过程中， 与病毒聚合酶结合。由于这些是病毒复制的重要步骤， 这两种方法都能有效地阻止病毒的传播。我们的基本工作表明，对于布尼亚病毒科， N的主要体外RNA底物是RNA柄状结构， 基因组末端负链病毒RNA的体内识别和双核苷化可能需要高水平的 N与锅柄的亲和相互作用。我们的方法将以这种特定的相互作用为基础 一种快速方便的检测方法来鉴定阻断N功能的分子。这很可能导致 鉴定干扰N与N的正确相互作用的新候选分子的星座， vRNA。将检查候选分子对细胞活力和病毒复制的影响。

The Bunyaviridae family of viruses include a set of Category A and C virus species. For the Bunyaviridae these include the members of the hantavirus genus, as well as Rift Valley fever virus and Crimean-Congo hemorrhagic fever virus. Current therapies against most of these viruses either do not exist or are only marginally efficient. Thus, it would be highly beneficial to identify and develop additional drugs directed at specific viral targets required for replication. The goal of this revised proposal is to refine and use an assay system for the identification of molecules that disrupt the function of the principle nucleocapsid protein of theses viruses, and to carry out screening for such candidate drugs. The nucleocapsid protein (N) functions both in the process of RNA encapsidation during virus assembly and during genome replication in conjunction with the viral polymerase. Since these are essential steps in virus replication, disruption of one or both processes would effectively block virus propagation. Our basic work indicates that for the Bunyaviridae, the principle in vitro RNA substrate for N is the RNA panhandle formed by the hydrogen bonding of the genome termini. In vivo discrimination and encapsidation of minus strand viral RNAs likely requires high affinity interaction of N with the panhandle. Our approach will be to use this specific interaction as the basis of a rapid and convenient assay to identify molecules that block N function. This is likely to lead to the identification of a constellation of novel candidate molecules that interfere the correct interaction of N with vRNA. Candidate molecules will be examined for their effect on cell viability and virus replication.