GENETICS OF 5 NONCODING REGIONS OF PICORNAVIRUS RNAS

小核病毒 RNA 5 个非编码区的遗传学

• 批准号: 2462852
• 负责人: Bert L Semler
• 金额: $ 24.27万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2462852
• 关键词: HeLa cells; Picornaviridae; RNA binding protein; RNA biosynthesis; gene mutation; genetic regulation; genetic regulatory element; genetic translation; nucleic acid sequence; nucleic acid structure; poliovirus; protein biosynthesis; protein sequence; ribonucleoproteins; ribosomes; site directed mutagenesis; tissue /cell culture; virus RNA; virus genetics; virus replication

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.

描述：申请人将继续他的研究，旨在了解 RNA-蛋白质相互作用在由5 '-NCR指导的功能中的作用 小核糖核酸病毒RNA 其他活动（如 病毒体组装）可以是 与小核糖核酸病毒基因组RNA的5 '-NCR相关，他将专注于他的 在功能上的努力（即， 病毒翻译和RNA复制）， 由这一区域的病毒RNA引导。 对于翻译功能， 测试假设，特定的核糖核蛋白复合物之间 在结合之前需要细胞蛋白和未加帽的病毒RNA 核糖体的启动帽非依赖性蛋白质合成。 为 复制功能，PI将测试假设， 在5 '-NCR中具有特定序列的细胞RNA结合蛋白（PCBP 2） 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 在5 '-NCR中含有定点损伤的突变体和假回复突变体 以及一些功能关联的生化分析 5 '-NCR与细胞或病毒蛋白之间的相互作用。 具体来说，PI 目的：（1）阐明神经细胞特异性因子在 脊髓灰质炎病毒翻译起始。 (2)确定细胞表达 PCBP 2的模式和生化特性。 (3)定义的功能 PCBP 2通过与茎环IV的相互作用参与帽非依赖性翻译 在脊髓灰质炎病毒RNA的IRES中。 (4)确定之间的关系 PCBP 2核糖核蛋白与茎环I和病毒RNA的复合物形成 复制的 申请人指出，这些方法，加上 分析突变病毒在HeLa细胞和神经元细胞中的生长特性， 应提供新的见解所采用的机制， 小核糖核酸病毒，以确保病毒的忠实翻译和复制 RNA在感染哺乳动物细胞过程中的作用。 他还表示， 从广义上讲，他的研究将导致一个高分辨率的“地图”的具体 RNA和蛋白质之间的大分子相互作用 影响细胞质中功能所需的高级结构 真核细胞