PROTEIN-FACILITATED RNA CATALYSIS

蛋白质促进的 RNA 催化

• 批准号: 6493847
• 负责人: Kevin M Weeks
• 金额: $ 2.91万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6493847
• 关键词: DNA footprinting; RNA binding protein; RNA biosynthesis; RNA splicing; covalent bond; crosslink; enzyme activity; enzyme complex; helicase; introns; nucleic acid sequence; protein structure function; ribonucleoproteins; ribozymes

DESCRIPTION (adapted from applicant's abstract): Dr. Kevin Weeks requests four years of support to elucidate fundamental features of how RNA and protein molecules collaborate to perform RNA catalysis. The proposal focuses on a mode ribonucleoprotein catalyst formed by the bI5 group I intron, a catalytic RNA, and the CBP2 protein. Catalysis provides a sensitive proof of structure-function relationships in RNA-protein interactions. This system is simple enough to be understood in significant detail, yet it is sophisticated enough to illustrate principles fundamental to more complex RNA-protein machines like those responsible for the biologically and medically important processes of RNA biogenesis, polypeptide synthesis, and chromosome maintenance Specific Aims are as follows. (1) Obtain a detailed three dimensional view of the specific bI5 structures involved in interaction with CBP2. A novel method of covalent footprinting for mapping direct RNA-protein interactions in comple RNAs will be developed. RNA sequence and structural requirements for protein-facilitated catalysis will be examined using iterative in vitro selection. This work will also address compromises involved in requiring proteins to facilitate RNA catalysis. (2) Determine specific functional roles for the two structural domains that comprise the CBP2 splicing factor and assign functions to individual domains in the context of a specific, detailed, and comprehensive kinetic framework for protein-facilitated RNA splicing. The assignment of individual protein domains to specific activities will promote understanding of the general roles of splicing factors in RNA catalysis. (3) Establish a mechanistic framework for understanding how Suv3 protein functions to disassemble the CBP2-RNA complex. Facilitated recycling of CBP2 is essentia for additional rounds of splicing on a biologically relevant time scale. These studies will define principles governing reorganization of sophisticated RNA-protein complexes by proteins whose function is coupled to NTP hydrolysis.

描述（根据申请人的摘要改编）：Kevin Weeks博士请求 四年的支持，以阐明RNA和RNA的基本特征 蛋白质分子合作执行RNA催化。 提案 专注于BI5组I形成的核糖核蛋白催化剂 内含子，催化RNA和CBP2蛋白。 催化提供了 RNA-蛋白质中结构功能关系的敏感证明 互动。 该系统足够简单，可以在重要的 细节，但它足够复杂，可以说明原理的基本原则 像更复杂的RNA蛋白机一样 生物学和医学上重要的RNA生物发生过程， 多肽合成和染色体维护特定目的是 跟随。 （1）获得特定BI5的详细三维视图 与CBP2相互作用涉及的结构。 一种新颖的共价方法 用于映射Comple RNA中直接RNA - 蛋白质相互作用的足迹将 被开发。 RNA序列和结构要求 将使用迭代体外检查蛋白质促催化的催化 选择。 这项工作还将解决涉及的妥协 蛋白质促进RNA催化。 （2）确定特定功能 构成CBP2剪接因子的两个结构域的角色 并在特定的上下文中将函数分配给单个域 针对蛋白质的RNA的详细且全面的动力学框架 剪接。 单个蛋白质结构域分配给特定 活动将促进对剪接一般角色的理解 RNA催化的因素。 （3）建立一个机械框架 了解SUV3蛋白如何拆卸CBP2-RNA 复杂的。 CBP2的促进回收利用是额外的回合 在生物学相关的时间范围内进行剪接。 这些研究将定义 控制复杂的RNA-蛋白质复合物的重组原则 通过其功能与NTP水解耦合的蛋白质。