Characterization of the reovirus simga 1s protien.

描述（由申请人提供）：了解病毒如何破坏正常的细胞过程，为了解宿主细胞功能的分子机制提供了巨大的见解。哺乳动物呼肠孤病毒感染是研究dsRNA病毒复制和发病机制的一种高度易处理的模型，它深刻地改变了宿主细胞生理，导致细胞周期阻滞和诱导凋亡，这一过程复杂地涉及癌症、免疫和发育。呼肠孤病毒非结构蛋白a1s与细胞周期的扰动和细胞凋亡的诱导有关。然而，a15介导这些作用的机制尚不清楚。利用一种新开发的基于质粒的呼肠孤病毒反向遗传系统，产生了含有野生型a1s编码S1基因片段或S1基因片段的病毒，其中a1s的翻译起始位点被改变以阻止a1s的合成。本应用程序中描述的实验将使用这些其他等基因病毒来定义a1s在呼肠孤病毒复制和发病机制中的功能，并确定a1s在呼肠孤病毒诱导的细胞周期阻滞和凋亡中的作用。提出了三个具体目标。第一个具体目标是确定a1s在体外培养细胞中呼肠孤病毒复制中的功能，并确定a1s是否有助于呼肠孤病毒诱导的细胞凋亡。第二个具体目标将确定a1s对细胞周期进程的影响，并使用蛋白质组学方法鉴定与a1s相互作用的细胞蛋白。第三个具体目标是确定a1s在呼肠孤病毒诱导的体内发病机制中的作用。总的来说，这些研究将为呼肠孤病毒感染调节宿主细胞周期和诱导细胞凋亡的机制提供新的见解。本应用中描述的实验目的是确定呼肠孤病毒a1蛋白在调节细胞周期进程、细胞凋亡和病毒发病机制中的功能。这些研究的发现可能会让我们对这些在发育、免疫和癌症中发挥重要作用的高度调控、相互关联的过程有新的认识。这些见解可能会导致对退行性、免疫或肿瘤疾病的新治疗。

DESCRIPTION (provided by applicant): Understanding how viruses disrupt normal cellular processes has provided tremendous insights into molecular mechanisms that underlie host cell function. Infection by mammalian reovirus, a highly tractable model for studies of dsRNA virus replication and pathogenesis, profoundly alters host cell physiology leading to cell cycle arrest and induction of apoptosis, processes intricately involved in cancer, immunity, and development. The reovirus nonstructural protein, a1s, is implicated in perturbation of the cell cycle and induction of apoptosis. However, mechanisms by which a1s mediates these effects are not known. Using a newly developed, plasmid-based reverse genetics system for reovirus, viruses were generated that contain either the wild-type a1s-encoding S1 gene segment or an S1 gene segment in which the a1s translational start site has been altered to block a1s synthesis. Experiments described in this application will use these otherwise isogenic viruses to define the function of a1s in reovirus replication and pathogenesis and determine the role of a1s in reovirus-induced cell cycle arrest and apoptosis. Three specific aims are proposed. The first specific aim will define the function of a1s in reovirus replication in cultured cells and determine whether a1s contributes to reovirus-induced apoptosis. The second specific aim will determine the effects of a1s on cell cycle progression and use proteomic approaches to identify cellular proteins with which a1s interacts. The third specific aim will define the contribution of a1s to reovirus-induced pathogenesis in vivo. Collectively, these studies will provide new insights into mechanisms by which reovirus infection modulates the host cell cycle and induces apoptosis. The goal of the experiments described in this application is to determine the function of reovirus a1s protein in modulating cell cycle progression, apoptosis, and viral pathogenesis. Findings from these studies could lead to new insights into these highly-regulated, interrelated processes that play fundamental roles in development, immunity, and cancer. Such insights might lead to new treatments for degenerative, immunologic, or neoplastic diseases.