Structural studies of RNase P

RNase P 的结构研究

• 批准号: 7903961
• 负责人: Alfonso Mondragon
• 金额: $ 27.8万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7903961
• 关键词: Address; Architecture; Area; Biochemical; Catalysis; Catalytic RNA; Cell physiology; Chemicals; Cleaved cell; Complex; Data; Development; Enzymes; Family; Funding; Goals; Holoenzymes; Indium; Ions; Knowledge; Lead; Life; Metals; Methods; Molecular Biology; Nucleotides; Organism; Process; Proteins; RNA; RNA Processing; RNase P; Reaction; Research Personnel; Resolution; Ribonucleases; Role; Site-Directed Mutagenesis; Structure; Thermotoga maritima; Transfer RNA; Work; X-Ray Crystallography; base; endonuclease; improved; interest; programs; tRNA Precursor; three dimensional structure

DESCRIPTION (provided by applicant): Transfer RNA is produced as a precursor molecule that needs to be processed both at its 3' and 5' ends. Ribonuclease P, or RNase P, is the only endonuclease responsible for processing the 5' end of tRNA by cleaving a precursor and leading to tRNA maturation. It is composed of a large RNA molecule and at least one protein and it has been identified in all organisms. It was one of the first catalytic RNA molecules discovered and its study has been pivotal to our understanding of the role of RNA molecules in catalysis. RNase P is a true multi-turnover ribozyme that recognizes its substrate in trans and one of only two universal ribozymes. The knowledge of the structure and function of RNase P promises to provide important and relevant information on a key ribozyme involved in a central cellular process common to all organisms and also to further our understanding of the structure and function of large RNA molecules. This proposal is concerned with the structure and function of RNase P. In the past few years we have made substantial progress, including solving the structure of the intact RNA component of Thermotoga maritima RNase P. This structure provided important structural information on the RNA component of RNase P and allowed us to make crucial observations regarding the function of this universal ribozyme and relate the structure to the wealth of existing biochemical data. For the next funding period we propose to continue and expand our studies of RNase P. The specific aims for this proposal are: 1) to determine the three dimensional structure of a complex of the RNase P holoenzyme with tRNA, 2) to extend our structural studies of the RNA component of T. maritima RNase P to higher resolution and, 3) to study the role of the universally conserved regions in the structure of RNase P and the implications for RNA recognition and cleavage. The work is based on a combination of molecular biology and biochemical methods to produce and characterize the molecules that we require for our work, and X-ray crystallography to solve their atomic structures.

描述（由申请人提供）：转移RNA是作为前体分子产生的，需要在其3‘和5’端进行处理。核糖核酸酶P，或RNase P，是唯一负责通过切割前体并导致tRNA成熟处理tRNA 5'端的内切酶。它由一个大的RNA分子和至少一种蛋白质组成，在所有生物体中都有发现。它是最早发现的催化RNA分子之一，它的研究对我们理解RNA分子在催化中的作用至关重要。RNase P是一种真正的多转换核酶，它识别反式底物，也是仅有的两种通用核酶之一。对RNase P结构和功能的了解有望为所有生物体共同参与中心细胞过程的关键核酶提供重要和相关的信息，也有助于我们进一步了解大RNA分子的结构和功能。在过去的几年里，我们已经取得了实质性的进展，包括解决了Thermotoga martima RNase P的完整RNA成分的结构，该结构提供了RNase P的RNA成分的重要结构信息，使我们能够对这种通用核酶的功能进行重要的观察，并将结构与现有的丰富的生化数据联系起来。未来融资期间我们建议继续和扩大我们的核糖核酸酶的研究P .这个提案的具体目标是:1)确定的三维结构复杂的核糖核酸酶P全酶的tRNA, 2)来扩展我们的结构性研究RNA组件的t . maritima核糖核酸酶P分辨率更高,3)研究普遍保守地区的角色对RNA核糖核酸酶P的结构和影响识别和乳沟。这项工作是基于分子生物学和生物化学方法的结合，以产生和表征我们工作所需的分子，以及x射线晶体学来解决它们的原子结构。