GENETICS OF 5' NONCODING REGIONS OF PICORNAVIRUS RNA'S

小核病毒 RNA 5 非编码区的遗传学

• 批准号: 6831187
• 负责人: Bert L Semler
• 金额: $ 33.56万
• 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 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6831187
• 关键词: GENETICS; NONCODING; REGIONS; PICORNAVIRUS; RNA

EXCEED THE SPACE PROVIDED. The experiments described in this proposal will address the translation and RNA replication functions encoded by the 5' noncoding region (5' NCR) of picornavirus genomic RNAs. These functions are the result of protein-protein, RNA-protein, and RNA-RNA interactions involving both viral and cellular components. The proposed experiments will address the roles of viral RNAs and cellular proteins as they pertain to the separate activities of translation initiation and RNA synthesis. However, because there is functional overlap between some of the components utilized during cap-independent translation initiation by picornaviruses encoding type I IRES elements and proteins involved in picornavirus RNA synthesis, some of the proposed experiments will converge on both processes. Experiments outlined in this proposal will define the domains in the cellular RNA-binding protein, poly(rC) binding protein (PCBP2), required for multimerization and/or RNA binding and how such domains contribute to viral functions in the cytoplasm of infected cells. Such experiments will address important mechanistic questions regarding the role of PCBP2 and other cellular polypeptides in picomavirus translation initiation. This proposal will also test a hypothesis posited to explain a putative switch between translation and RNA replication on the same template (or templates) during a picornavirus infectious cycle. Finally, in vivo cell culture experiments are proposed that utilize recently- described experimental approaches to modulate protein levels and domain functions of cellular proteins hypothesized to participate in specific steps in the viral life cycle that involve the genomic RNA 5' NCR. Results from these proposed studies will provide new mechanistic insights into the complex processes involved in picornavirus translation initiation and into the interplay between translation functions and viral RNA replication. In addition, data generated from such studies should reveal fundamental aspects of RNA- protein interactions relevant to cellular cap-independent translation as well as pre-mRNA splicing and other elements of RNA metabolism. PERFORMANCE SITE ========================================Section End===========================================

超出所提供的空间。本提案中描述的实验将解决小核糖核酸病毒基因组 RNA 的 5' 非编码区 (5' NCR) 编码的翻译和 RNA 复制功能。这些功能是涉及病毒和细胞成分的蛋白质-蛋白质、RNA-蛋白质和RNA-RNA相互作用的结果。拟议的实验将探讨病毒 RNA 和细胞蛋白的作用，因为它们涉及翻译起始和 RNA 合成的单独活动。然而，由于编码 I 型 IRES 元件的小核糖核酸病毒在帽独立翻译起始过程中使用的一些组件与参与小核糖核酸病毒 RNA 合成的蛋白质之间存在功能重叠，因此一些拟议的实验将集中在这两个过程上。该提案中概述的实验将定义多聚化和/或RNA结合所需的细胞RNA结合蛋白、聚(rC)结合蛋白(PCBP2)中的结构域，以及这些结构域如何促进受感染细胞细胞质中的病毒功能。此类实验将解决有关 PCBP2 和其他细胞多肽在小核糖核酸病毒翻译起始中的作用的重要机制问题。该提案还将测试一个假设，该假设旨在解释小核糖核酸病毒感染周期期间同一模板（或多个模板）上翻译和 RNA 复制之间的假定转换。最后，提出了体内细胞培养实验，利用最近描述的实验方法来调节假设参与病毒生命周期中涉及基因组RNA 5'NCR的特定步骤的细胞蛋白的蛋白质水平和结构域功能。这些拟议研究的结果将为小核糖核酸病毒翻译起始所涉及的复杂过程以及翻译功能和病毒RNA复制之间的相互作用提供新的机制见解。此外，此类研究产生的数据应该揭示与细胞帽独立翻译相关的RNA-蛋白质相互作用的基本方面以及前mRNA剪接和RNA代谢的其他要素。表演网站==========================================章节结束===============================================