Mechanistic Studies on Sindbis Virus Replication

正义RNA病毒代表了一组不同的经济和医学上重要的病原体。这些病毒的基因组RNA是一种多功能分子，作为病毒蛋白翻译的mRNA和合成负义RNA复制中间体的模板。为了确定基因组功能调控的手段，本项目重点研究Sindbis病毒（甲病毒属的类型种）由正义基因组RNA编程并由病毒复制酶完成的负链RNA合成。这项研究的长期目标是建立一个Sindbis病毒基因表达和基因组复制的综合模型。本项目将研究Sindbis病毒基因组5‘和3’端的RNA序列元件如何影响病毒复制酶对模板的识别，调节模板功能，并促进负链RNA合成的有效起始。利用实验室建立的实验系统，该研究旨在：1)绘制基因组5‘区域的决定因子及其同源结合因子，以合成负链RNA； 2)阐明基因组3’端的残基促进负链RNA合成的有效起始的机制。这些研究的结果将特别影响对甲病毒基因表达和复制的理解。此外，由于单链正义RNA病毒是由一个共同的基因组复制方案联合起来的，从这些研究中建立的原理将对这类病毒具有更广泛的意义。该项目将包括培养研究生和一名博士后研究员。此外，来自该大学生命科学领域代表性不足群体项目的学生将参与该研究项目。

Positive-sense RNA viruses represent a diverse group of economically and medically important pathogens. The genomic RNA of these viruses is a multi-functional molecule that serves as an mRNA for translation of virus proteins and a template for the synthesis of a minus-sense RNA replication intermediate. In order to determine the means by which genome function is regulated this project focuses on Sindbis virus (the type species of the genus Alphavirus) minus-strand RNA synthesis programmed by the positive-sense genomic RNA and committed by the viral replicase. The long-term goal of the research is to develop an integrated model of Sindbis virus gene expression and genome replication. This project will investigate how RNA sequence elements at the 5' and 3' end of the Sindbis virus genome effect template recognition by the viral replicase, regulate template function, and facilitate productive initiation of minus-strand RNA synthesis. Using experimental systems established in the laboratory the research aims to: 1) map determinants in the 5' region of the genome and their cognate binding factors required for minus-strand RNA synthesis, and 2) elucidate the mechanism by which residues at the 3' end of the genome facilitate productive initiation of minus-strand RNA synthesis. Results from these studies will specifically impact the understanding of alphavirus gene expression and replication. Further, as single-stranded positive-sense RNA viruses are united by a common scheme of genome replication, principles established from these studies will have broader implications for this group of viruses generally. The project will involve the training of graduate students and a postdoctoral researcher. In addition, students from the university's program for underrepresented groups in the life sciences will be participants in this research project.