Molecular Mechanisms in Reovirus mRNA Synthesis

呼肠孤病毒 mRNA 合成的分子机制

• 批准号: 6709385
• 负责人: MAX L. NIBERT
• 金额: $ 34.4万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-10 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6709385
• 关键词: RNA binding protein; RNA biosynthesis; Reoviridae; cryoelectron microscopy; genetic transcription; intracellular transport; messenger RNA; posttranscriptional RNA processing; protein structure function; recombinant proteins; virus RNA; virus assembly; virus genetics; virus protein

DESCRIPTION (provided by applicant): The molecular mechanisms by which mammalian orthoreoviruses (reoviruses) and other dsRNA viruses mediate synthesis of their mRNA molecules using virally encoded enzymes packaged within infectious virus particles is a subject of active inquiry because they promise insight into how the steps in mRNA synthesis - plus-strand RNA synthesis (transcription), RNA 5' capping, and RNA transport - occur within the delimited three-dimensional setting of the icosahedral virus particle. A better understanding of these mechanisms in dsRNA viruses should be useful for understanding how related processes are mediated by analogous other viral, microbial, or cellular enzymes and for designing new antiviral, antimicrobial, or anticellular agents directed at them. The long-term objective of our work with reoviruses is to define structure-function relationships for the roles of reovirus proteins in viral replication and effects on host cells and animals. In the current proposal, emphasis is placed on the proteins within the reovirus core (subviral) particle that mediates mRNA synthesis in vitro. Three specific aims are identified, which reflect where further progress appears most promising or necessary to advance understanding in this system. These aims are (1) to determine the structure of the reovirus transcriptase complexes and their arrangement in cores, (2) to define the assembly pathway of the reovirus core shell, and (3) to dissect the functions of the reovirus core proteins in RNA synthesis, capping, and transport. Reovirus particles reconstituted from recombinant proteins expressed using baculovirus vectors play a prominent role in the proposed experiments because of their broad applicability to studies of particle structure, assembly, and functions. Recently determined crystal structures of the transcription- and capping-competent reovirus core particle and the reovirus RNA-dependent RNA polymerase also figure heavily in this proposal by having focused attention on particular areas where more structure information is needed, suggested specific new hypotheses about steps in mRNA synthesis and assembly, and identified specific amino acids in the core proteins to subject to mutagenesis for structure-function testing. The results of these studies will enhance our understanding of the reovirus core as an elegantly designed molecular machine for mRNA synthesis.

描述（由申请人提供）：分子机制 哺乳动物矫形病毒（蠕虫病毒）和其他DSRNA病毒介导 使用包装的病毒编码的酶合成其mRNA分子 传染性病毒颗粒是主动查询的主题，因为它们承诺 深入了解mRNA合成中的步骤如何 - 加链RNA合成 （转录），RNA 5'上限和RNA传输 - 出现在划界中 二维病毒颗粒的三维设置。更好 了解DSRNA病毒中的这些机制应该对 了解相关过程是如何通过类似的其他病毒介导的， 微生物或细胞酶，用于设计新的抗病毒抗菌剂， 或针对它们的抗细胞剂。我们工作的长期目标 与静脉病毒一起定义结构功能关系的关系 病毒复制中的葡萄病毒蛋白对宿主细胞和动物的影响。 在当前的提案中，重点放在依默病毒中的蛋白质上 在体外介导mRNA合成的核心（次病）粒子。三个具体 确定了目标，这反映了进一步进展最大的位置 有望或必要的，以提高该系统中的理解。这些目标是 （1）确定葡萄病毒转录酶复合物的结构和 它们在核心中的排列，（2）定义依孢病毒的组装途径 核心外壳，（3）剖析依克肠疾病的功能 RNA合成，封盖和运输。葡萄病毒颗粒从 使用杆状病毒载体表达的重组蛋白起着重要作用 在拟议的实验中，由于它们对研究的广泛适用性 粒子结构，组装和功能。最近确定的晶体 转录和限额的依默病毒核心粒子的结构 以及依赖RNA RNA聚合酶RNA的依赖病毒也很大程度上构成 提案通过将注意力集中在更多结构的特定领域 需要信息，建议的有关mRNA步骤的特定新假设 合成和组装，并确定核心中的特定氨基酸 蛋白质以诱变进行结构功能测试。结果 在这些研究中，将增强我们对依孢病毒核心的理解 优雅设计的分子机，用于mRNA合成。