RNA PROTEIN INTERACTIONS IN POLIOVIRUS REPLICATION

描述（改编自申请人的总结）：本项目的长期目标 项目旨在了解脊髓灰质炎病毒复制的机制及其 与宿主细胞的相互作用。 具体而言，申请人有兴趣 研究RNA合成的起始及其与翻译的关系。 重点是解剖核糖核蛋白的结构和功能 复合物（RNP-B），似乎参与启动 正链RNA合成。 核糖核酸这种核糖核蛋白的核糖核酸成分 一个复合物折叠成苜蓿叶状结构， 3CD和核糖体相关因子（p36/PCBP）。 的身份 最近已经确定了宿主细胞因子，但是 核糖核蛋白复合物尚未被定义。 安迪诺博士发明了一种 研究脊髓灰质炎病毒复制的新系统。 微量注射 脊髓灰质炎病毒RNA进入非洲爪蟾卵母细胞启动了一个完整的， 真实的病毒复制周期，产生高水平的传染性 病毒，但只有当脊髓灰质炎病毒RNA与存在于 HeLa细胞 两种HeLa细胞因子是病毒复制所必需的。 卵母细胞，一个参与翻译的起始，另一个参与RNA 合成. 因此，卵母细胞中的显微注射基本上可以用作一种免疫抑制剂。 鉴定和进一步分析病毒功能的体外系统 和细胞因子，并从生物化学的角度剖析 引发脊髓灰质炎病毒RNA合成。 本提案的具体目的是确定 NRP-B复合物，研究其在卵母细胞和人类细胞中的功能， 确定脊髓灰质炎病毒在卵母细胞中复制所需的人为因素。

DESCRIPTION (Adapted from Applicant's Summary): The long-term goal of this project is to understand the mechanism of poliovirus replication and its interaction with the host-cell. Specifically, the applicant is interested in studying initiation of RNA synthesis and its relationship to translation. The focus is on dissecting the structure and function of a ribonucleoprotein complex (RNP-B) that appears to participate in the initiation of positive-strand RNA synthesis. The RNA component of this ribonucleoprotein complex folds into a cloverleaf-like structure and binds the viral protein 3CD and a ribosome-associated factor (p 36/PCBP). The identity of the host-cell factor has recently been determined, but the function of the ribonucleoprotein complex has yet to be defined. Dr. Andino has developed a novel system in which to study poliovirus replication. Microinjection of poliovirus RNA into Xenopus laevis oocytes initiates a complete and authentic viral replication cycle, which yields a high level of infectious viruses, but only if polioviral RNA is co-infected with factors present in HeLa cells. Two HeLa cell factors are required for viral replication in oocytes, one involved in initiation of translation, and the other in RNA synthesis. Thus, microinjection in oocytes can be used essentially as an in vitro system in which to identify and further analyze the function of viral and cellular factors, and to biochemically dissect the mechanism of initiation of poliovirus RNA synthesis. The specific aims of this proposal are to determine the structure of the NRP-B complex, to study its function in oocytes and human cells, and to characterize human factors require for poliovirus replication in oocytes.