HOST-PATHOGEN INTERACTION IN VIRAL REPLICATION

内科科学家的职业发展需要一个广泛的方法来为学术医学的研究、教学和临床责任做准备。这项资助提出了一个为期四年的职业发展计划，主要包括有针对性的假设驱动型研究，但也包括教学和指导活动，参与机构和国家会议，以及持续的临床培训。威斯康星大学麦迪逊分校为医生和科学家提供了理想的职业发展环境，以一流的医疗保健和尖端的生物医学研究而闻名，拥有最先进的设施，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。本研究将探讨病毒复制和发病机制中涉及的动态和复杂的宿主-病原体相互作用。正链RNA病毒的简单基因组结构和实验证据表明，宿主因子对其复制是不可或缺的，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然未知。最近在已经被充分研究过的酵母菌中新病毒复制系统的发展，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。禽舍病毒（FHV）是一种双部正链RNA病毒，是noddaviridae家族的成员，是唯一一种在葡萄球菌中复制的动物病毒，因此代表了一种独特的模型来研究病毒复制机制。酿酒酵母实验系统将用于分离和表征不能支持FHV复制的酵母突变体。同时免疫化学方法也将用于分离和表征与FHV RNA聚合酶相关的细胞宿主蛋白。宿主-病原体相互作用的鉴定和表征，特别强调细胞宿主因子的影响，对于了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都至关重要。

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.