HOST-PATHOGEN INTERACTION IN VIRAL REPLICATION

医生-科学家的职业发展需要广泛的方法来准备在学术医学的职业生涯中承担研究、教学和临床责任。这笔赠款提出了一项为期四年的职业发展计划，主要包括有针对性的假设驱动的研究，但也包括教学和指导活动，参加机构和国家会议，以及继续临床培训。威斯康星大学麦迪逊分校以一流的医疗保健和尖端的生物医学研究、最先进的设施、广泛的技术和智力资源以及研究人员和临床医生之间开放合作的强大传统而闻名，为发展内科科学家的职业生涯提供了理想的环境。这项拟议的研究将调查涉及病毒复制和发病机制的动态和复杂的宿主-病原体相互作用。正链RNA病毒简单的基因组结构和实验证据表明，宿主因子对于它们的复制是必不可少的，尽管这些宿主因子的身份及其对病毒复制的功能影响在很大程度上仍不清楚。最近在研究得很好的酿酒酵母中开发了新的病毒复制系统，这使得对参与正链RNA病毒复制的细胞宿主因子的功能研究成为可能。鸡群屋病毒(FHV)是一种双链正链RNA病毒，属于诺达病毒科，是唯一在酿酒酵母中复制的动物病毒，是研究病毒复制机制的独特模型。酿酒酵母实验系统将用于分离和鉴定无法支持FHV复制的酵母突变株。同时还将使用一种免疫化学方法来分离和鉴定与FHV RNA聚合酶相关的细胞宿主蛋白。宿主-病原体相互作用的识别和表征，特别是细胞宿主因素的影响，对于理解病毒复制和发病的基本机制以及设计合理的抗病毒治疗方法都是至关重要的。

The career development of physician-scientists requires a broad approach in preparation for a career in academic medicine with research, teaching, and clinical responsibilities. This grant proposes a four-year career development plan consisting primarily of targeted hypothesis-driven research, but also incorporating teaching and mentoring activities, participation in institutional and national conferences and meetings, and continued clinical training. The University of Wisconsin-Madison provides the ideal environment to develop a career as a physician-scientist, with a reputation for premiere medical care and cutting-edge biomedical research, state-of-the-art facilities, extensive technical and intellectual resources, and a strong tradition of open collaboration among researchers and clinicians. The proposed research will investigate the dynamic and complex host-pathogen interactions involved in viral replication and pathogenesis. The simple genomic structure of positive-strand RNA viruses and experimental evidence suggest that host factors are indispensable for their replication, although the identity of these host factors and their functional impact on viral replication remain largely unknown. The recent development of novel viral replication systems in the well-studied yeast Saccharomyces cerevisiae permits the functional investigation of the cellular host factors involved in positive-strand RNA virus replication. Flock house virus (FHV), a bipartite positive-strand RNA virus and member of the Nodaviridae family, is the only animal virus shown to replicate in S. cerevisiae, and consequently represents a unique model to investigate the mechanisms of viral replication. The S. cerevisiae experimental system will be used to isolate and characterize yeast mutants unable to support FHV replication. A concurrent immunochemical approach will also be used to isolate and characterize cellular host proteins associated with the FHV RNA polymerase. The identification and characterization of host-pathogen interactions, with a particular emphasis on the impact of cellular host factors, are crucial both to understand the basic mechanisms of viral replication and pathogenesis and to design rational antiviral therapies.