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。对于布尼亚病毒科 这些病毒包括汉坦病毒属的成员，以及裂谷热病毒和克里米亚-刚果 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 略有效率。因此，确定和开发针对以下目标的额外药物将是非常有益的 复制所需的特定病毒靶点。这个修订提案的目标是改进和使用一种化验方法 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，并对这些候选药物进行筛选。核衣壳蛋白(N)的功能 在病毒组装和基因组复制期间的RNA封装过程中 与病毒聚合酶结合。由于这些是病毒复制的基本步骤，因此中断一个或多个 这两个过程都将有效地阻止病毒传播。我们的基本工作表明，对于布尼亚病毒科来说， N的体外RNA底物的原理是由N的氢键形成的RNA狭长柄 基因组末端。在体内识别和包裹负链病毒RNA可能需要很高的 N与狭长柄的亲和力作用。我们的方法将是使用这种特定的交互作为基础 一种快速而方便的方法来识别阻断N功能的分子。这很可能导致 识别干扰N与N的正确相互作用的新的候选分子星座 VRNA。候选分子将被检测它们对细胞活性和病毒复制的影响。

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.