Structure and Dynamics of RNA

RNA的结构和动力学

• 批准号: 6652577
• 负责人: ARTHUR PARDI
• 金额: $ 25.56万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6652577
• 关键词: RNA; chemical structure function; conformation; fluorescence resonance energy transfer; gene mutation; nuclear magnetic resonance spectroscopy; nucleic acid structure; spectrometry; transfer RNA

DESCRIPTION (provided by applicant): This proposal will continue our program on NMR studies of RNA structure and dynamics. Although new biological functions for RNAs continue to be discovered, there is still relatively little information on the three-dimensional structures of RNAs, as compared with proteins. One component of this project involves development of improved NMR methods for solution structure determinations of RNA, including application of TROSY experiments for studies of larger molecules. We will also develop improved methods for incorporating residual dipolar couplings into solution structure determinations of RNA. Residual dipolar coupling data provide long-range angle information that complements standard short-range distance and torsion angle NMR data. This long-range structural information makes it possible to dramatically improve the global structure determination of nucleic acids in solution. Methods will also be developed for obtaining additional dipolar couplings constraints and improving the RNA structure refinements with residual dipolar coupling data.The residual dipolar coupling methods for determining global structures of molecules will be applied to a series of tRNA and tRNA-like molecules. We will study the global conformation of a number of standard and unusual cytosolic and mitochondrial tRNAs. Some of these mitochondrial tRNA mutants have been implicated in various human diseases, and we will determine if mutations that cause disease lead to changes in the global structure of the tRNA.The dynamics of a common RNA tertiary structural motif, the GAAA tetraloop-receptor interaction, will be studied by 13C relaxation and residual dipolar coupling NMR experiments. The relaxation experiments probe the local dynamics, and the dipolar couplings probe domain dynamics for the RNA. Recently developed single molecule fluorescence resonance energy transfer techniques will also be used to study dynamics for helical domains in this GAAA-tetraloop tertiary motif. This combination of NMR relaxation, NMR dipolar coupling and single molecule fluorescence techniques will provide a comprehensive picture of the conformational fluctuations over a wide range of timescales. These data will lead to a better understanding of how dynamics correlates with the function and stability of this RNA tertiary structural motif.

描述（由申请人提供）：该提案将继续我们的 RNA 结构和动力学 NMR 研究项目。尽管RNA新的生物学功能不断被发现，但与蛋白质相比，关于RNA三维结构的信息仍然相对较少。该项目的一个组成部分涉及开发改进的 NMR 方法来测定 RNA 的溶液结构，包括应用 TROSY 实验研究较大分子。我们还将开发改进的方法，将残余偶极耦合纳入 RNA 溶液结构测定中。残余偶极耦合数据提供长程角度信息，补充标准短程距离和扭转角 NMR 数据。这种长程结构信息使得显着改善溶液中核酸的整体结构测定成为可能。还将开发用于获得额外偶极耦合约束并利用残余偶极耦合数据改进RNA结构细化的方法。用于确定分子整体结构的残余偶极耦合方法将应用于一系列tRNA和类tRNA分子。我们将研究许多标准和不寻常的胞质和线粒体 tRNA 的整体构象。其中一些线粒体 tRNA 突变体与各种人类疾病有关，我们将确定导致疾病的突变是否会导致 tRNA 整体结构的变化。常见 RNA 三级结构基序（GAAA 四环受体相互作用）的动态，将通过 13C 弛豫和残余偶极耦合 NMR 实验进行研究。弛豫实验探测局部动力学，偶极耦合探测 RNA 的域动力学。最近开发的单分子荧光共振能量转移技术也将用于研究 GAAA-四环三级基序中螺旋结构域的动力学。 NMR 弛豫、NMR 偶极耦合和单分子荧光技术的结合将提供广泛时间尺度内构象波动的全面图像。这些数据将有助于更好地理解动力学如何与该 RNA 三级结构基序的功能和稳定性相关。