Hantavirus RNA Encapsidation

这是一个修订的应用程序，用于检查新诺布雷病毒(SNV)复制过程中RNA包裹的机制。SNV是一种A类病原体，属于布尼亚病毒科汉坦病毒属。我们预计，在SNV感染过程中发生的RNA包膜的一般原理将适用于汉坦病毒属的其他成员，并可能适用于整个布尼亚病毒科。自提交原始提案以来，我们获得了大量额外的初步数据。这些初步数据让我们得出两个主要假设。首先，病毒末端的狭长柄RNA结构是病毒核衣壳蛋白识别的顺式作用包膜序列。其次，核衣壳蛋白是一种非特异性的RNA伴侣，是产生具有生物功能的高阶病毒RNA结构所必需的。这项R21应用中提出的工作将表征核衣壳与病毒RNA狭长柄之间的相互作用，并研究核衣壳蛋白的RNA伴侣活性的作用机制。我们预计， SNV的病毒RNA和核衣壳蛋白的一般性质适用于汉坦病毒属的其他成员，也适用于布尼亚病毒科的其他成员。 我们实验室以前的工作主要集中在慢病毒的封装、RNA结构以及封装二聚体HIV RNA所需的顺式和反式组件上。我们过去对这种正链RNA病毒的经验可能会对我们研究负链汉坦病毒RNA的包裹体有用。我们已经生成了一些与汉坦病毒RNA封装相关的方面的初步数据，其中一些数据在本提案中进行了描述。然而，由于我们最近才开始使用汉坦病毒系统，还没有为我们拟议的工作提供资金，因此申请R21似乎是合适的。在我们的初步调查之后，我们的较长期计划包括试图为SNV和布尼亚病毒科的其他成员生成一幅全面的RNA包裹图，并寻求 通过RO1机制提供资金。除了表征汉坦病毒封装的生物学特征外，我们的长期目标还将包括尝试通过设计这些病毒的筛选策略来利用这些信息，并可能通过为各种应用生成基于汉坦病毒的载体。

This is a revised application to examine the mechanism of RNA encapsidation during replication of Sin nombre virus (SNV). SNV is a Category A pathogen and a member of the hantavirus genus in the Bunyaviridae family. We anticipate that the general principles of RNA encapsidation that take place during SNV infection will be applicable to the other members of the hantavirus genus and likely to the entire Bunyaviridae family. Since submission of the original proposal we have obtained substantial additional preliminary data. These preliminary data lead us to two major hypotheses. First, the terminal viral panhandle RNA structure is the cis-acting encapsidation sequence recognized by the viral nucleocapsid protein. Second, the nucleocapsid protein is a nonspecific RNA chaperone required for the generation of biologically functional higher order viral RNA structure. The work proposed in this R21 application will characterize the interaction between the nucleocapsid and the viral RNA panhandle, and also examine the mechanism of action of the RNA chaperone activity of the nucleocapsid protein. We anticipate that the general properties of the viral RNA and nucleocapsid protein of SNV be applicable to other members of the hantavirus genus and also to other members of the Bunyaviridae family. Previous work from our lab focused on lentivirus encapsidation, RNA structure, and the cis- and transcomponents required for encapsidation of dimeric HIV RNA. Our past experience with this positive stranded RNA virus will likely be useful as we investigate encapsidation of negative stranded hantavirus RNA. We have generated some preliminary data concerning facets relevant to hantavirus RNA encapsidation, and some of these data are described in this proposal. However, since we have only recently embarked on the hantavirus system and do not yet have funding for our proposed work, it seemed appropriate to apply for an R21. Following our preliminary investigations, our longer range plans include trying to generate a comprehensive picture of RNA encapsidation for SNV, and other members of the Bunyaviridae, and to seek funding through the RO1 mechanism. In addition to characterizing the features of the biology of hantavirus encapsidation, our longer term goals will also include attempting to make use of this information by devising screening strategies for these viruses and perhaps by generating hantavirus-based vectors for various applications.