Structural studies of the RNA component of RNase P

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'末端进行加工。核糖核酸酶RNase是tRNA P或P的5'端的内切核酸酶，仅负责通过切割前体并导致tRNA成熟的加工。它含有一种RNA和一种蛋白质成分，已在所有生物体中鉴定。它是最早被发现的催化RNA分子之一，它的研究对于我们理解RNA分子在催化中的作用至关重要。核糖核酸酶P是一种真正的多周转核酶，也是仅有的两种在所有三个生命王国中保守的核酶之一。核糖核酸酶P的结构和功能的知识，承诺提供重要的和相关的信息，一个关键的核酶参与一个中央的细胞过程中常见的所有生物体，也进一步了解大RNA分子的结构和功能。细菌RNase P的RNA组分由两个结构域组成：特异性结构域和催化结构域。最近，我们解决了B的特异性结构域的结构。枯草杆菌RNA酶P，我们现在建议扩大我们的研究，以更充分地表征B的特异性结构域。枯草杆菌RNase P，以研究其他细菌RNase P的结构，并研究完整全酶的结构。本文的具体目标是：1）确定T. 2）获得关于B的更高分辨率信息。3）研究B.枯草杆菌RNase P与金属离子及其底物，和4）结晶和溶解完整的B结构。枯草RNA酶P全酶。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结构。

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 RNase is the endonuclease for the 5' end of tRNA P, or P, only responsible processing by cleaving a precursor and leading to tRNA maturation. It contains an RNA and a protein component and has been identified in all organisms. It is one of the first catalytic RNA molecules identified 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 and one of only two ribozymes conserved in all three kingdoms of life. 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 bacterial RNase P. The RNA component of bacterial RNase P is made of two domains: a specificity and a catalytic domain. Recently, we solved the structure of the specificity domain of B. subtilis RNase P and we are now proposing to expand our studies to characterize more fully the specificity domain of B. subtilis RNase P, to study the structure of other bacterial RNase P, and to study the structure of the intact holoenzyme. The specific aims for this proposal are: 1) to determine the structure of the T. thermophilus RNase P specificity domain, 2) to obtain higher resolution information on the B. subtilis RNase P specificity domain, 3) to study the interactions of B. subtilis RNase P with metal ions and with its substrate, and 4) to crystallize and solve the structure of the intact B. subtilis RNase P holoenzyme. 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 structures.