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Structure of RNA and RNP

RNA 和 RNP 的结构

基本信息

批准号：
14035205
负责人：
YOKOYAMA Shigeyuki
金额：
$ 47.17万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14035205/
关键词：
X-ray Crystallography NMR 3D structure RNA Intermolecular interaction RNA-binding protein アミノアシルtRNA合成酵素 LINE RNA修飾酵素 tmRNA SRP RNAポリメラーゼリボゾーム RNAポリメーゼ

项目摘要

X-ray crystallography: By crystallography of RNA-protein complexes, structural details of their interactions and roles of RNA in activities were analyzed. We determined crystal structures of aminoacyl-tRNA synthetases, which are responsible for joining corresponding tRNAs and amino acids during translation. It revealed that tRNAs play positive roles in both aminoacylation and proofreading reactions by modulating protein structures. To analyze structural bases of pre-tRNA maturation, structural studies have been done on enzymes responsible for the pre-tRNA processing and modifications. The structure of tRNA complexed with ArcTGT revealed that the modification is achieved via a step including a transitional tRNA structure "λ form". We also determined the crystal structure of Vasa, an RNA helicase involved in the differentiation of germ cells, and elucidated structural basis of the RNA double strand resolution by a DEAD-box RNA helicase.NMR: (1) Solution structures of helix 6 stem-loop RNA of SRP RNA from human and Pyrococcus furiosus were determined and the difference in recognition by SRP19 was discussed. (2) Solution structure of SmpB from Thermus thermophilus HB8 was determined and the interaction between tmRNA and SmpB was analyzed. It was found that the structure of SmpB changed upon binding of tmRNA. (3) Solution structures of stem-loop RNAs derived from UnaL2 RNA were determined. The GNRNA motif in the penta loop was found to be required for the recognition by the LINE reverse transcriptase and the flexibility of the bulged stem was found to be important for the retrotransposition initiation reaction. (4) A classification system for RNA structures, CSNA, was developed and applied for RNA structure determination by NMR.

X射线晶体学：通过对RNA-蛋白质复合物的晶体学研究，分析了它们相互作用的结构细节以及RNA在活性中的作用。我们确定了氨酰-tRNA合成酶的晶体结构，这些合成酶负责在翻译过程中连接相应的tRNA和氨基酸。结果表明，tRNA通过调节蛋白质结构，在氨酰化反应和校对反应中发挥积极作用。为了分析前体tRNA成熟的结构基础，已经对负责前体tRNA加工和修饰的酶进行了结构研究。与Arc TGT复合的tRNA的结构表明，修饰是通过包括过渡tRNA结构“λ形式”的步骤实现的。我们还测定了与生殖细胞分化有关的RNA解旋酶Vasa的晶体结构，阐明了DEAD-box RNA解旋酶拆分RNA双链的结构基础。NMR：（1）测定了人和激烈火球菌SRP RNA的6螺旋茎环RNA的溶液结构，并讨论了SRP 19对6螺旋茎环RNA的识别差异。(2)测定了嗜热栖热菌（Thermus thermophilus）HB 8的SmpB的溶液结构，分析了SmpB与tmRNA的相互作用。结果发现，SmpB的结构发生变化时，结合的tmRNA。(3)测定了来源于UnaL 2 RNA的茎环RNA的溶液结构。发现五环中的GNRNA基序是LINE逆转录酶识别所需的，并且发现凸出茎的柔性对于逆转录转座起始反应是重要的。(4)建立了一个RNA结构分类系统CSNA，并将其应用于NMR测定RNA结构。