GENETICS OF 5 NONCODING REGIONS OF PICORNAVIRUS RNAS

描述：申请人将继续他的学习，目的是了解 RNA-蛋白质相互作用在5‘-NCR调控的功能中的作用 小核糖核酸。虽然其他活动(如病毒粒子组件)可以 与小核糖核酸基因组RNA的5‘-NCR相关，他将把他的 在功能上的努力(即病毒翻译和RNA复制)已知 是由病毒RNA的这个区域引导的。对于翻译职能，他将 验证以下假设：特定的核糖核蛋白复合体 在结合之前需要细胞蛋白和未封顶的病毒RNA 核糖体，用于启动帽非依赖性蛋白质合成。对于 复制函数，PI将检验假设 5‘-NCR中具有特定序列的细胞RNA结合蛋白(PCBP2) 脊髓灰质炎病毒RNA和病毒蛋白3CD是启动 病毒RNA合成。他将利用他收集的可行的 5‘-NCR中含有定点损伤的突变体和假逆转株 以及一些用于功能关联的生化分析 在5‘-NCR和细胞或病毒蛋白之间。具体地说，PI 将：(1)阐明神经细胞特异性因子在 脊髓灰质炎病毒翻译启动。(2)确定细胞表达 PCBP2的型式和生化性质。(3)明确功能定位 PCBP2与茎环IV相互作用在帽非依赖性翻译中的作用 在脊髓灰质炎病毒RNA的IRES中。(四)确定相互之间的关系 与茎环I和病毒RNA形成PCBP2核糖核蛋白复合体 复制。申请人说，这些方法，再加上 突变病毒在HeLa细胞和神经细胞中的生长特性分析， 应该会提供对 微小核糖核酸病毒确保病毒的忠实翻译和复制 在哺乳动物细胞感染过程中的RNA。他还表示，更多 总的来说，他的研究将导致一幅高分辨率的特定地图 核糖核酸与蛋白质在蛋白质编配中的大分子相互作用 在细胞质中影响功能所需的更高级结构 真核细胞。

DESCRIPTION: The applicant will continue his studies aimed at understanding the roles of RNA-protein interactions in functions directed by the 5'-NCR of picornavirus RNAs. Although other activities (eg. virion assembly) may be associated with the 5'-NCR of picornavirus genomic RNAs, he will focus his efforts on functions (ie. viral translation and RNA replication) known to be directed by this region of viral RNAs. For translation functions he will test the hypothesis that specific ribonucleoprotein complexes between cellular proteins and uncapped viral RNAs are required prior to binding ribosomes for initiation of cap-independent protein synthesis. For the replication functions, the PI will test the hypothesis that interaction of a cellular RNA binding protein (PCBP2) with a specific sequence in the 5'-NCR of poliovirus RNA and the viral protein 3CD is necessary for initiation of viral RNA synthesis. He will take advantage of his collection of viable mutants and pseudorevertants containing site-directed lesions in the 5'-NCR as well as a number of biochemical assays for functional associations between the 5'-NCR and cellular or viral proteins. Specifically, the PI will: (1) Elucidate the role of neuronal-cell specific factors in poliovirus translation initiation. (2) Determine the cellular expression pattern and biochemical properties of PCBP2. (3) Define the function of PCBP2 in cap-independent translation via its interaction with stem-loop IV in the IRES of poliovirus RNAs. (4) Determine the relationship between PCBP2 ribonucleoprotein complex formation with stem-loop I and viral RNA replication. The applicant states that these approaches, coupled with the analysis of mutant viral growth properties in HeLa cells and neuronal cells, should provide new insights into the mechanisms employed by the picornaviruses to insure the faithful translation and replication of viral RNAs during an infection of mammalian cells. He also indicated that, more broadly, his studies will lead to a high resolution "map" of specific macromolecular interactions between RNAs and protein in the orchestration of higher order structures required for effecting functions in the cytoplasm of eukaryotic cells.