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计划，核磁共振研究的RNA结构和动力学。尽管RNA的新的生物学功能不断被发现，但与蛋白质相比，关于RNA的三维结构的信息仍然相对较少。该项目的一个组成部分是开发用于RNA溶液结构测定的改进NMR方法，包括应用TROSY实验研究更大的分子。我们还将开发改进的方法，将残余偶极耦合到RNA的溶液结构测定。残余偶极耦合数据提供了补充标准短程距离和扭转角NMR数据的长程角度信息。这种远程结构信息使得有可能显着改善溶液中核酸的全局结构测定。我们还将发展一些方法来获得额外的偶极耦合约束，并利用剩余偶极耦合数据来改进RNA结构的精细化。用于确定分子全局结构的剩余偶极耦合方法将应用于一系列tRNA和tRNA样分子。我们将研究一些标准的和不寻常的胞质和线粒体tRNA的全球构象。这些线粒体tRNA突变体中的一些已被牵连在各种人类疾病，我们将确定是否突变，导致疾病导致的全球结构的变化的tRNA.The一个共同的RNA三级结构基序，GAAA四环受体相互作用的动力学，将通过13 C弛豫和残余偶极耦合NMR实验研究。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在这个GAAA-tetraloop三级基序的螺旋结构域的动力学。这种NMR弛豫，NMR偶极耦合和单分子荧光技术的组合将提供一个全面的图片的构象波动在很宽的时间范围内。这些数据将导致更好地理解动力学如何与这种RNA三级结构基序的功能和稳定性相关。