Structure and Dynamics of RNA

RNA的结构和动力学

• 批准号: 6545052
• 负责人: ARTHUR PARDI
• 金额: $ 28.87万
• 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/6545052
• 关键词: RNA; chemical structure function; conformation; fluorescence resonance energy transfer; 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）：该提案将继续我们对RNA结构和动力学的NMR研究计划。尽管继续发现RNA的新生物学功能，但与蛋白质相比，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涉及开发改进的NMR方法来确定RNA的溶液结构，包括将Trosy实验应用于较大分子的研究。我们还将开发改进的方法，将残留的偶极耦合纳入RNA的溶液结构确定。残留的偶极耦合数据提供了远程角度信息，可补充标准的短距离距离和扭转角度NMR数据。这种远程结构信息使得有可能显着改善溶液中核酸的整体结构测定。还将开发用于获得其他偶极耦合约束的方法，并使用残留的偶性偶联数据改进RNA结构的改进。用于确定分子全球结构的残留偶极耦合方法将应用于一系列TRNA和TRNA类似分子。我们将研究许多标准和不寻常的胞质和线粒体TRNA的全球构象。这些线粒体tRNA突变体中的一些已与各种人类疾病有关，我们将确定引起疾病的突变是否会导致tRNA的全球结构发生变化。共同的RNA三级结构基序的动力学，GAAA TETRALOOP-TETRALOOP-RECTECTOR将通过13C弛豫和持续的二色性Dipalal dipalal coupling nmreents进行研究。松弛实验探测了局部动力学，以及RNA的偶极耦合探针域动力学。最近开发的单分子荧光共振能量传递技术也将用于研究此GAAA-TETRALOOP三级基序中的螺旋结构域的动力学。 NMR松弛，NMR偶极耦合和单分子荧光技术的这种结合将为众多范围的构象波动提供全面的图像。这些数据将更好地理解动力学与该RNA三级结构基序的功能和稳定性的相关性。