MOLECULAR MECHANISMS IN REOVIRUS TRANSCRIPTION & CAPPING

拟议的研究将提供关于如何 哺乳动物呼肠孤病毒的转录和封端酶被组织起来 在亚病毒“核心”粒子内。一般的方法包括 5种核心蛋白(Gamma1、Gamma2、Gamma3、MU2和 Sigma2)，用于研究它们的结构、功能和组装成 粒子与更经典的遗传和生物化学研究相关联 病毒在转录、封端和输出过程中的不同活性 具有特定核心蛋白及其结构域的信使RNA。长的- 这项工作的术语目标是在全分子、结构 以及机械感知呼肠孤病毒核心如何调节这些活动。这个 这些发现可以应用于理解和控制这些过程 病毒和其他病原体，以及在患病和正常的真核生物中 细胞。该项目的具体目标是：(1)确定 所有五种核心蛋白的结构位置和结构域组织， (2)研究核糖核酸依赖的核苷三磷酸酶活性。 并确定其在转录或封端中的作用，(3) 关联四种封闭酶活性(RNA三磷酸酶、RNA 鸟苷酸转移酶、RNA[鸟氨酸-7-N]-甲基转移酶和RNA[核糖- 2‘-O]-甲基转移酶)中含有特定的蛋白质或蛋白质区域 核心，以及(4)确定基因组dsRNA片段是如何组织的和 与核心中的蛋白质相互作用。

The proposed studies will provide fundamental information about how the transcription and capping enzymes of mammalian reoviruses are organized within the subviral "core" particle. The general approach involves expression of the five core proteins (gamma1, gamma2, gamma3, mu2, and sigma2) for studies of their structure, function, and assembly into particles and more classical genetic and biochemical studies to associate the different activities in transcription, capping, and export of the viral messenger RNAs with specific core proteins and their domains. The long- term goal of this work is to characterize in a full molecular, structural, and mechanical sense how reovirus cores mediate these activities. The findings can be applied to understanding and controlling these processes in viruses and other pathogens, as well as in diseased and normal eukaryotic cells. The specific aims of the project are (1) to determine the structural positions and domain organizations of all five core proteins, (2) to investigate the RNA-dependent nucleoside triphosphatase activity in cores and to identify its role in transcription or capping, (3) to affiliate the four capping enzyme activities (RNA triphosphatase, RNA guanylyltransferase, RNA [guanine-7-N]-methyltransferase, and RNA [ribose- 2'-O]-methyltransferase) with specific proteins or protein regions in the core, and (4) to determine how the genomic dsRNA segments are organized and interact with proteins in the core.