Mechanistic Studies on Sindbis Virus Replication

辛德毕斯病毒复制机制研究

• 批准号: 0749482
• 负责人: Richard Hardy
• 金额: $ 45万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-15 至 2012-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0749482&HistoricalAwards=false
• 关键词: Mechanistic; Studies; Sindbis; Virus; Replication

Virus replication is achieved through the precise coordination of a series of macromolecular synthetic events. For plus-strand RNA viruses the temporal regulation of gene expression and genome replication occurs through the action of the viral ribonuclear protein complex (vRNP). The vRNP is assembled around viral RNA and its composition is defined by the recruitment of proteins, both viral and host, to specific RNA elements. Differential availability of proteins and viral RNA elements at particular times during the virus replication cycle can therefore alter the composition of the vRNP and change the function of the complex. Identifying RNA elements required for specific macromolecular synthetic events and the proteins that bind them to form a vRNP are fundamental requirements if we are to understand the regulatory processes necessary for virus replication.This project seeks to understand the regulation of gene expression and genome replication of plus-strand RNA viruses using Sindbis virus (SIN), the type species of the alphavirus genus. During SIN replication the genome is translated giving rise to the non-structural polyprotein. A semi-processed form of this protein is required for the copying (minus-strand synthesis) of the genome from which it was translated. In turn the minus-strand RNA serves as a template for production of progeny genomes, and a subgenomic mRNA synthesized by a complex containing the fully processed non-structural proteins. It is therefore apparent that different vRNP complexes perform each of these processes. The project aims to elucidate the molecular interactions that mediate the transitions in vRNP composition and function These studies will significantly impact the understanding of the molecular mechanisms required for alphavirus replication. Additionally, as plus-strand RNA viruses are united by a common scheme of replication, principles established from these studies will be of more general interest and importance for the study of all plus-strand RNA viruses. The project will provide a forum for the training of undergraduate students, graduate students, and post-doctoral fellows. A number of students from various programs (including Initiative for Maximizing Student Diversity Program and Research Experience for Undergraduates Program) have participated in research in the laboratory and it is fully expected that this participation will continue.

病毒复制是通过一系列大分子合成事件的精确协调来实现的。对于正链RNA病毒，基因表达和基因组复制的时间调节通过病毒核糖核蛋白复合物（vRNP）的作用发生。vRNP围绕病毒RNA组装，其组成由病毒和宿主蛋白质募集至特定RNA元件来定义。因此，在病毒复制周期的特定时间，蛋白质和病毒RNA元件的差异可用性可以改变vRNP的组成并改变复合物的功能。要想了解病毒复制的调控过程，首先要确定特定大分子合成所需的RNA元件和与之结合形成vRNP的蛋白质。本课题以甲病毒属的模式种--辛德毕斯病毒（SIN）为对象，研究正链RNA病毒的基因表达和基因组复制的调控。在SIN复制过程中，基因组被翻译，产生非结构多蛋白。这种蛋白质的半加工形式是复制（负链合成）它所翻译的基因组所必需的。反过来，负链RNA作为产生后代基因组的模板，以及由含有完全加工的非结构蛋白的复合物合成的亚基因组mRNA。因此，很明显，不同的vRNP复合物执行这些过程中的每一个。该项目旨在阐明介导vRNP组成和功能转变的分子相互作用。这些研究将显著影响对甲病毒复制所需分子机制的理解。此外，由于正链RNA病毒通过共同的复制方案结合在一起，因此从这些研究中建立的原理对于所有正链RNA病毒的研究将具有更普遍的意义和重要性。该项目将为本科生、研究生和博士后研究员的培训提供一个论坛。来自不同项目（包括最大限度地提高学生多样性计划和本科生研究经验计划）的一些学生参加了实验室的研究，预计这种参与将继续下去。