STRUCTURE OF VIRAL RNA AND ITS ROLE IN ASSEMBLY

病毒RNA的结构及其在组装中的作用

• 批准号: 6788734
• 负责人: ALEXANDER MCPHERSON
• 金额: $ 30.33万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6788734
• 关键词: Alphaherpesvirinae; Bromovirus; RNA virus; X ray crystallography; atomic force microscopy; capsid; computer simulation; conformation; human immunodeficiency virus; intermolecular interaction; murine leukemia virus; nephelometry; nucleic acid structure; tobacco mosaic virus; vaccinia virus; virus RNA; virus assembly

DESCRIPTION (provided by applicant): A variety of physical and chemical techniques, chief among them being X-ray crystallography, will be applied to a series of icosahedral viruses with the ultimate objectives of (1) deducing the detailed structure of the RNA genomes inside the viruses, (2) identifying the important protein-nucleic acid interactions, and (3) devising models for the assembly and disassembly of the virions consistent with existing molecular biological data. Viruses to be studied are Satellite Tobacco Mosaic Virus (STMV), Satellite Tobacco Necrosis Virus (STNV), Satellite Panicum Mosaic Virus (SPMV), Turnip Yellow Mosaic Virus (TYMV), Desmodium Yellow Mottle Virus (DYMV), Tomato Aspermy Virus (TAV), Brome Mosaic Virus (BMV), Panicum Mosaic Virus (PMV), and empty capsids of each of the last five species. These are viruses whose structures are best known, for which RNA is at least partially visible in the virion, and for which the greatest amount of experience exists. Core RNA particles will be prepared by digestion of the intact viruses and will be crystallized for X-ray diffraction analyses. Empty capsids will be made, crystallized, and their diffraction data used in difference Fourier syntheses to reveal RNA structure. With integration of the proposed chemical and enzymatic probe information, and predictions from computer modeling to supplement the X-ray studies, a comprehensive reconciliation of structure with function will emerge. We have shown that when atomic force microscopy is applied in conjunction with chemical and enzymatic reagents, complex viruses can be dissected to reveal internal components, including nucleic acid. To extend our investigations of nucleic acid conformation and packaging to larger, more complex viruses we will extend our AFM investigations of the internal structures of HIV, herpes simplex virus, mouse leukemic virus, and vaccinia virus. This research is significant because the conformation of the nucleic acid component of no virus is currently known, nor is there any comprehensive structural basis for the processes of viral assembly and disassembly.

描述（由申请人提供）：各种物理和化学技术，其中主要是X射线晶体学，将应用于一系列二十面体病毒，最终目标是（1）推断病毒内部RNA基因组的详细结构，（2）识别重要的蛋白质-核酸相互作用，以及（3）设计与现有分子一致的病毒体组装和分解模型 生物数据。要研究的病毒有卫星烟草花叶病毒（STMV）、卫星烟草坏死病毒（STNV）、卫星黍花叶病毒（SPMV）、萝卜黄花叶病毒（TYMV）、金钱草黄斑驳病毒（DYMV）、番茄不育病毒（TAV）、雀麦花叶病毒（BMV）、 黍花叶病毒 (PMV)，以及最后五个物种的空衣壳。这些病毒的结构最为人所知，RNA在病毒颗粒中至少部分可见，并且存在最丰富的经验。核心 RNA 颗粒将通过消化完整病毒来制备，并结晶用于 X 射线衍射分析。将制作空衣壳并使其结晶，并将其衍射数据用于差异傅里叶合成以揭示 RNA 结构。通过整合所提出的化学和酶探针信息，以及计算机建模的预测来补充 X 射线研究，将出现结构与功能的全面协调。我们已经证明，当原子力显微镜与化学和酶试剂结合应用时，可以解剖复杂的病毒以揭示内部成分，包括核酸。为了将我们对核酸构象和包装的研究扩展到更大、更复杂的病毒，我们将扩大对 HIV、单纯疱疹病毒、小鼠白血病病毒和牛痘病毒内部结构的 AFM 研究。这项研究意义重大，因为目前尚不清楚病毒核酸成分的构象，也没有病毒组装和分解过程的任何全面的结构基础。