NUCLEIC ACID INTERACTIONS IN FLOCK HOUSE VIRUS

鸡群病毒中的核酸相互作用

• 批准号: 6012902
• 负责人: NEEL KUMAR KRISHNA
• 金额: $ 3.17万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6012902
• 关键词: RNA virus; X ray crystallography; computer simulation; dimer; electron microscopy; insect virus; intermolecular interaction; nucleic acid denaturation; nucleic acid structure; virion; virus RNA; virus assembly

The experiments outlined in this proposal focus on RNA-RNA interactions in Flock House virus (FHV), a nonenveloped, icosahedral insect virus. The genome of FHV is split between two distinct, single-stranded, messenger-sense RNA molecules which are thought to be encapsidated into a single virion. The mechanism by which one copy of each genomic RNA is selected for packaging into a single particle is unknown. Preliminary studies from our laboratory have demonstrated that mild heating of purified virions induces formation of a stable heterodimeric RNA complex within the particle suggesting that the encapsidated nucleic acids undergo significant structural rearrangements. Furthermore, mild dissociation of FHV provirions has revealed that this complex may already exist prior to heating, albeit in a more labile form. Heterodimer formation is hypothesized to provide a means by which the two RNAs are copackaged during assembly of virus particles. In this application, we intend to isolate the labile heterodimer from purified virions and/or reconstitute it in vitro and plan to characterize it using various biochemical, physical and molecular genetic methods. In addition, in collaboration with Dr. John E. Johnson's laboratory, a newly developed low resolution X-ray crystallography technique will be utilized to visualize changes in RNA structure induced upon heating of FHV particles. The results from the proposed studies should provide new insights into the mechanisms of RNA packaging in segmented viruses and the organization of the nucleic acid inside the virion. This knowledge could provide a basis for developing strategies of how to interfere with the life cycle of medically important viruses.

该提案中概述的实验重点介绍了羊群病毒（FHV）中的RNA-RNA相互作用，这是一种未开发的二十面体昆虫病毒。 FHV的基因组分为两个不同的单链，信素的RNA分子，这些分子被认为被封装成单个病毒体。 每个基因组RNA的一个副本被选择用于包装到单个粒子中的机制尚不清楚。 我们实验室的初步研究表明，纯化病毒体的轻度加热诱导粒子中稳定的异二聚体RNA复合物的形成，这表明封装的核酸经历了显着的结构重排。此外，FHV规定的轻度解离表明，尽管以更不稳定的形式，该复合物可能已经存在于加热之前。 假设杂化二聚体形成提供了一种在病毒颗粒组装过程中两个RNA共同包装的手段。 在此应用中，我们打算将不稳定的异二聚体与纯化的病毒体分离，并在体外重新构造它，并计划使用各种生化，物理和分子遗传学方法对其进行表征。 此外，与John E. Johnson博士的实验室合作，将利用一种新开发的低分辨率X射线晶体学技术可视化FHV颗粒加热后引起的RNA结构的变化。 拟议的研究的结果应提供有关分段病毒中RNA包装机制以及病毒座内部核酸的组织的新见解。 这些知识可以为制定如何干扰医学上重要病毒的生命周期的策略提供基础。