Hantavirus RNA Encapsidation and Packaging

描述（由申请人提供）：这是一份修订后的申请，旨在全面检查汉坦病毒核衣壳蛋白（N）在病毒复制中的作用。汉坦病毒包括A类病原体，并且是布尼亚病毒科（Bunyaviridae）的成员，布尼亚病毒科包括A类和C类病原体。汉他病毒感染期间发生的RNA包装原理很可能适用于汉他病毒属的其他成员，整个布尼亚病毒科，以及其他医学上重要的分段负链RNA病毒，如流感病毒。所有负链、节段RNA病毒都具有通过基因组末端的相互作用形成“锅柄”的基因组片段。我们已经发现，这种病毒RNA（vRNA）柄是汉坦病毒N的主要高亲和力结合底物。我们将进行互补的体外和体内实验，以表征病毒RNA正确包装成颗粒所需的决定因素。我们还发现汉坦病毒N蛋白是一种强有力的RNA分子伴侣，它能瞬时解离RNA的高级结构，促进具有生物学功能的高级结构的形成。因此，我们将进行实验来表征N-相关RNA伴侣蛋白功能在病毒复制中的作用。特别是，我们将描述的作用，N在锅柄形成和启动病毒RNA复制。我们还发现，N模拟翻译起始期间帽结合复合物的多个细胞肽的活性。我们将进一步描述N在这个过程中的作用。虽然N在复制中的多个步骤中发挥作用，但我们认为N的共同内在活动介导了这些生物学功能。除了表征汉坦病毒N的生物学特征外，我们还将利用我们观察到的N和vRNA柄之间的高亲和力相互作用来鉴定抑制N功能和病毒复制的分子。 公共卫生相关性：汉坦病毒引起汉坦病毒肺综合征，在人类中具有高死亡率。与汉坦病毒密切相关的其他病毒引起多种人类疾病，包括出血热。我们提出的工作的目标是确定病毒复制的关键蛋白之一的突出的生物学功能，并确定药物有效地阻止这些病毒的复制。

DESCRIPTION (provided by applicant): This is a revised application to comprehensively examine the roles of hantavirus nucleocapsid protein (N) in virus replication. Hantaviruses include Category A pathogens and are members of the Bunyaviridae family which includes both Category A and C pathogens. It is likely that the principles of RNA packing that take place during hantavirus infection will be applicable to the other members of the hantavirus genus the entire Bunyaviridae family, and other medically important segmented minus strand RNA viruses such as influenza. All minus strand, segmented RNA viruses have genome segments that are found in "panhandle" formation via interaction of the genome termini. We have found that this viral RNA (vRNA) panhandle is the primary high affinity binding substrate for hantavirus N. We will carry out complementary in vitro and in vivo experiments to characterize the determinants required for correct packaging of viral RNA into particles. We have also found that hantavirus N protein is a robust RNA chaperone that transiently dissociates higher order RNA structure to facilitate formation of higher order structures with biological function. Thus, we will carry out experiments to characterize the role of N- associated RNA chaperone function in virus replication. In particular we will characterize the role of N in panhandle formation and in the initiation of viral RNA replication. We have also found that N mimics the activity of multiple cellular peptides of the cap-binding complex during translation initiation. We will further characterize the role of N in this process. Although N functions in multiple steps in replication, we think that common intrinsic activities of N mediate these biological functions. In addition to characterizing the features of the biology of hantavirus N, we will use our observed high affinity interaction between N and the vRNA panhandle to identify molecules that inhibit N function and virus replication. PUBLIC HEALTH RELEVANCE: Hantaviruses cause hantavirus pulmonary syndrome which has a high mortality rate in humans. Other viruses closely related to hantaviruses cause a diverse set of human diseases including hemorrhagic fever. The goal of our proposed work is to identify salient biological functions of one of the viral proteins crucial to virus replication and to identify drugs effective in blocking the replication of these viruses.