ARBOVIRUS REPLICATION IN MOSQUITO AND VERTEBRATE CELLS

这个项目的总体目标仍然是了解这些属性如何 节肢动物传播的病毒使它们能够以截然不同的方式复制 并在它们的脊椎动物宿主中引起各种不同的疾病 各州。 我们的实验系统将涉及辛德比斯和登革热的研究。 病毒(甲型和黄病毒属：Togaviridae科)以及 水泡性口炎病毒(横纹病毒科)在脊椎动物和 蚊子细胞。预计我们在世界各地了解到的这些病毒 细胞水平最终将增强我们对发生在 整个有机体，脊椎动物或无脊椎动物。在A.与VSV合作。 白纹伊蚊细胞，我们将研究病毒RNA合成的速度是如何 与蛋白质合成的抑制有关，最终与细胞杀伤有关。 导致蛋白质合成抑制的生化损伤将 被调查。 节肢动物传播的病毒，由于其广泛的宿主范围，是突出的 适合于寄主范围突变体的产生和研究。我们将寻求 额外的寄主范围突变体，并试图从分子方面确定 为什么这些突变体的复制能力受到限制 单元类型。我们还将确定核苷酸序列的变化，这些变化 与寄主范围表型相关。 辛德比斯病毒在白纹伊蚊细胞中的高效复制 被核苷类似物利巴韦林抑制。另一方面，这一点 在一定条件下，化合物对病毒的作用很小或没有。 在BHK细胞中复制。我们将继续确认这些财产 决定利巴韦林和WE疗效的不同宿主细胞 应评估其在整个动物不同器官中的抗病毒作用。 关于登革热病毒的复制策略和 一般的黄病毒比已知的甲型病毒或 弹状病毒。在这个项目中，我们打算研究蛋白质的模式 不同病毒特异性蛋白的加工及其功能 被感染的细胞。特别是对特定蛋白质的加工将是 与病毒的形态发生和成熟有关。

The overall goal of this project remains to understand how the properties of arthropod-borne viruses enable them to replicate in widely different hosts and to cause in their vertebrate hosts a variety of different disease states. Our experimental systems will involve the study of Sindbis and dengue viruses (genera alpha- and flavivirus: Family Togaviridae) as well as vesicular stomatitis virus (Family Rhabdoviridae) in vertebrate and mosquito cells. It is expected that what we learn of these viruses at the cellular level will eventually enhance our understanding of what occurs in the whole organism, vertebrate or invertebrate. Working with VSV in A. albopictus cells, we shall study how the rate of viral RNA synthesis is related to inhibiton of protein synthesis and eventually to cell killing. The biochemical lesion which leads to inhibition of protein synthesis will be investigated. Arthropod-borne viruses, because of their wide host range, are eminently suited for the generation and study of host-range mutants. We shall seek additional host-range mutants and attempt to determine in molecular terms why these mutants are restricted in their ability to replicate in a given cell type. We shall also determine nucleotide sequence changes which are specifically associated with the host-range phenotype. The replication of Sindbis virus in A. albopictus cells is efficiently inhibited by the nucleoside analog, ribavirin. On the other hand, this compound under certain conditions has little or no effect on viral replication in BHK cells. We shall continue to identify those properties of different host cells which determine the efficacy of ribavirin and we shall assess its anti-viral action in different organs of the whole animal. Much less is known of the strategy of replication of dengue virus and flaviviruses in general than is known about either alphaviruses or rhabdoviruses. In this project we intend to study the patterns of protein processing and the functions of the different virus-specified proteins in infected cells. In particular the processing of specific proteins will be related to viral morphogenesis and maturation.