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的感染是研究dsRNA病毒复制和发病机制的高度易处理的模型，它深刻地改变了宿主细胞生理学，导致细胞周期停滞和诱导细胞凋亡，这些过程复杂地参与癌症、免疫和发育。呼肠孤病毒的非结构蛋白a1s参与细胞周期的扰动和细胞凋亡的诱导。然而，a1s介导这些效应的机制尚不清楚。使用新开发的基于质粒的呼肠孤病毒反向遗传学系统，产生含有野生型a1s编码S1基因片段或其中a1s翻译起始位点已被改变以阻断a1s合成的S1基因片段的病毒。在本申请中描述的实验将使用这些在其他方面同基因的病毒来定义a1在呼肠孤病毒复制和发病机制中的功能，并确定a1在呼肠孤病毒诱导的细胞周期停滞和细胞凋亡中的作用。提出了三个具体目标。第一个具体的目标将确定在培养细胞中的呼肠孤病毒复制的功能，并确定是否a1s有助于呼肠孤病毒诱导的细胞凋亡。第二个具体目标是确定a1对细胞周期进程的影响，并使用蛋白质组学方法来鉴定与a1相互作用的细胞蛋白。第三个具体的目标是确定α 1对体内呼肠孤病毒诱导的发病机制的贡献。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导致对这些高度调节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。这些见解可能会导致新的治疗退行性疾病，免疫性疾病或肿瘤疾病。

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.