Characterization of the reovirus simga 1s protien.

描述(由申请人提供)：了解病毒如何扰乱正常的细胞过程，为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个高度容易处理的模型，它深刻地改变了宿主细胞的生理，导致细胞周期停滞和诱导细胞凋亡，这些过程复杂地参与了癌症、免疫和发育。呼肠孤病毒的非结构蛋白A1S与细胞周期紊乱和诱导细胞凋亡有关。然而，A1调节这些效应的机制尚不清楚。利用一种新开发的基于质粒的呼肠孤病毒反向遗传学系统，产生了含有野生型A1 S编码S1基因片段或S1基因片段的病毒，其中A1 S翻译起始点已被改变以阻止A1 S的合成。本申请中描述的实验将使用这些其他同源病毒来确定A1S在呼肠孤病毒复制和发病中的功能，并确定A1S在呼肠孤病毒诱导的细胞周期停滞和凋亡中的作用。提出了三个具体目标。第一个特定目标将确定A1在培养细胞中呼肠孤病毒复制中的功能，并确定A1是否参与呼肠孤病毒诱导的细胞凋亡。第二个特定目标将确定A1对细胞周期进程的影响，并使用蛋白质组学方法来鉴定A1与之相互作用的细胞蛋白质。第三个特定目标将定义A1在呼肠孤病毒诱导的体内致病中的作用。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。本申请中描述的实验的目的是确定呼肠孤病毒A1S蛋白在调节细胞周期进展、细胞凋亡和病毒致病中的功能。这些研究的发现可能导致对这些高度调控的、相互关联的过程的新见解，这些过程在发育、免疫和癌症中发挥着重要作用。这样的见解可能会为退行性、免疫性或肿瘤性疾病带来新的治疗方法。

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.