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作为前体分子产生，需要在其3'和5'末端处理。核糖核酸酶P或RNase P是唯一通过裂解前体并导致tRNA成熟来处理tRNA的5'端的核酸内切酶。它由一个大的RNA分子和至少一种蛋白质组成，并且在所有生物体中都被鉴定出来。它是发现的第一个催化RNA分子之一，其研究对于我们对RNA分子在催化中的作用的理解至关重要。 RNase P是一种真正的多旋转核酶，它识别其在反式的底物，而仅有两个通用核酶之一。 RNase P的结构和功能的知识有望提供有关所有生物共有的中心细胞过程的关键核酶的重要信息，也可以进一步了解我们对大RNA分子的结构和功能的理解。该建议与RNase P的结构和功能有关。在过去的几年中，我们取得了重大进展，包括解决Thermotoga maritima rnase的完整RNA成分的结构。这种结构提供了有关RNase P的RNA成分的重要结构信息，并使我们能够对这种通用核酶的功能进行重要的观察，并使其与该结构相关联。在下一个资金期间，我们建议继续并扩大对RNase P的研究。该提案的具体目的是：1）确定RNase P Holoenzyme与tRNA的复合物的三维结构，2）扩展我们对T. Maritima rnase P到更高分辨率和3）角色的结构的结构性研究，以研究和研究的作用。 RNA识别和乳沟。这项工作基于分子生物学和生化方法的结合，以产生和表征我们为工作所需的分子以及X射线晶体学以求解其原子结构。