Mechanism of RVFV nucleocapsid targeting drugs

描述（由申请方提供）：裂谷热病毒（RVFV）是一种蚊媒新发传染性病原体，可能对人类和农业动物健康产生重大影响。它是CDC/USDA A类选择因子，并已被世界动物卫生组织认定为具有全球传播潜力的高后果病原体。RVFV感染没有经过证实的治疗方法，也没有被证明安全有效的许可疫苗。我们最近在一种新的分子靶点高通量筛选中鉴定了78种化合物，它们干扰RVFV核衣壳蛋白N和靶RNA的相互作用。这些化合物来自现有的药物和药物样分子文库，并且几种化合物显示出对细胞培养物中RVFV复制的可测量的抑制。我们建议确定几个最有前途的化合物的分子作用机制。我们将使用生物化学、生物物理学和遗传学方法来确定化合物在N上的结合位点，并且我们将确定它们是否抑制特异性RNA结合活性或一般RNA结合活性， 都是N所固有的。具体而言，我们将确定抑制性化合物是否直接干扰RNA结合，干扰核衣壳蛋白寡聚化，或具有不同于RNA-蛋白或蛋白-蛋白界面的靶标，使用RNA和野生型和突变核衣壳蛋白的药物结合测定。这项研究通过扩大我们对病毒生长和抑制的理解，以及通过在对抗RVFV感染的新药物疗法方面取得进展，有望为生物医学和农业科学领域带来好处。

DESCRIPTION (provided by applicant): Rift Valley fever virus (RVFV) is a mosquito-borne emerging infectious agent with potential for major impacts on human and agricultural animal health. It is a CDC/USDA Category A Select Agent, and has been recognized by the World Organization for Animal Health as a high consequence pathogen with potential for global spread. There are no proven treatments for RVFV infections and there is no licensed vaccine that has been proven safe and effective. We recently identified 78 compounds in a novel molecular target high throughput screen that interfere with the interaction of RVFV nucleocapsid protein, N, and a target RNA. These compounds come from existing drug and drug-like molecule libraries and several of the compounds show measurable inhibition of RVFV replication in cell culture. We propose to ascertain the molecular mechanism of action of several of the most promising compounds. We will use biochemical, biophysical and genetic approaches to determine the binding site(s) for the compounds on N and we will determine whether they inhibit the specific RNA binding activity or the generic RNA binding activity that are both intrinsic to N. Specifically, we will determine whether the inhibitory compounds interfere directly with RNA binding, interfere with nucleocapsid protein oligomerization, or have a target that is distinct from either the RNA-protein or protein-protein interfaces using RNA and drug binding assays with wild-type and mutant nucleocapsid protein. The research promises benefits to the fields of biomedicine and agricultural science by expanding our understanding of virus growth and inhibition, and by making advances toward new drug therapies to combat RVFV infections.