Proteins in RNA-Based Enzymes

基于 RNA 的酶中的蛋白质

基本信息

批准号：
7848372
负责人：
ANDREY S. KRASILNIKOV
金额：
$ 28.57万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7848372
关键词：
Affect Binding Sites Biochemical Biochemistry Biological Assay Biological Process Catalysis Catalytic RNA Cell Cycle Regulation Cell Death Complex Development Disease Dwarfism Enzymes Eukaryota Eukaryotic Cell Evolution Family Goals Hair Holoenzymes Human Immunologic Deficiency Syndromes In Vitro Knowledge Lead Light Location Messenger RNA Modeling Mutation Nuclear Nucleotides Organism Play Process Protein Binding Protein Subunits Proteins Public Health RNA RNA Processing RNA Splicing RNA-Protein Interaction RNAse MRP RNase P Reliance Ribonucleases Ribonucleoproteins Ribosomal RNA Role Saccharomyces cerevisiae Specificity Therapeutic Agents Time Transfer RNA Translations X-Ray Crystallography Yeasts base cartilage development enzyme activity human disease in vivo mRNA Transcript Degradation novel therapeutics numb protein protein function research study structural biology

项目摘要

DESCRIPTION (provided by applicant): RNA-based enzymes are crucial in many important biological processes including translation and RNA processing. While in some RNA-based enzymes the RNA moiety alone is capable of performing catalysis, many of them contain proteins that are required for their function. Our proposal focuses on nuclear Ribonuclease MRP (RNase MRP), which is an excellent model for understanding the interplay of proteins and RNA in ribonucleoprotein complexes. RNase MRP is a eukaryotic ribonucleoprotein complex involved in the maturation of rRNA and the regulation of the cell cycle in yeast. RNase MRP is closely related to RNase P and is highly conserved in eukaryotes. S. cerevisiae RNase MRP contains ten proteins and a 340 nucleotide long RNA component. The similarity between RNase MRP and RNase P strongly suggests that catalysis in RNase MRP is RNA-based; however, as in the case of RNase P, all of RNase MRP proteins are also required for activity in vivo. It is particularly intriguing that this reliance on proteins in the enzymes of the RNase P/MRP family tends to grow in more evolutionary advanced organisms. Little is known about why the enzymes of the RNase P/MRP family acquired more and more proteins as evolution progressed, or why RNase MRP needs proteins when the catalytic RNA moiety seems to be essentially preserved. This lack of knowledge represents a problem since it hinders further understanding of these important enzymes. This proposal focuses on biochemical and structural studies of S. cerevisiae nucleolar RNase MRP and aims to understand how the interactions between the components of this ribonucleoprotein complex affect its activity and specificity. The specific aims of the proposal are: 1. To determine how protein subunits of RNase MRP interact with the RNA moiety; 2. To determine roles of RNase MRP protein components in the enzyme's activity and specificity. We will characterize the RNA-protein interactions in RNase MRP using RNA footprinting, mutational studies, and structural analysis, and determine roles of the protein moiety in the activity of RNase MRP. These experiments will clarify the functions of the proteins in RNase MRP and related enzymes. Relevance to Public Health RNase MRP is an RNA-based enzyme which is required for the survival of eukaryotic cells. Mutations in the RNA component of RNase MRP in humans cause the development of Cartilage Hair Hypoplasia, a severe multi-systemic disorder resulting in dwarfism and immunodeficiency, and a study of RNase MRP has a potential to shed a light on the cause of the disease.

描述（由申请人提供）：基于 RNA 的酶在许多重要的生物过程中至关重要，包括翻译和 RNA 加工。虽然在一些基于 RNA 的酶中，RNA 部分本身就能够发挥催化作用，但其中许多酶都含有其功能所需的蛋白质。我们的提案重点关注核核糖核酸酶 MRP (RNase MRP)，它是了解核糖核蛋白复合物中蛋白质和 RNA 相互作用的绝佳模型。 RNase MRP 是一种真核核糖核蛋白复合物，参与 rRNA 的成熟和酵母细胞周期的调节。 RNase MRP 与 RNase P 密切相关，在真核生物中高度保守。酿酒酵母 RNase MRP 含有 10 种蛋白质和 340 个核苷酸长的 RNA 成分。 RNase MRP 和 RNase P 之间的相似性强烈表明 RNase MRP 的催化作用是基于 RNA 的；然而，与 RNase P 一样，所有 RNase MRP 蛋白也是体内活性所必需的。特别有趣的是，这种对 RNase P/MRP 家族酶中蛋白质的依赖往往在进化程度更高的生物体中增长。人们对为什么 RNase P/MRP 家族的酶随着进化的进展获得越来越多的蛋白质，或者为什么当催化 RNA 部分似乎基本上被保留时 RNase MRP 需要蛋白质，知之甚少。这种知识的缺乏是一个问题，因为它阻碍了对这些重要酶的进一步了解。该提案重点关注酿酒酵母核仁 RNase MRP 的生化和结构研究，旨在了解这种核糖核蛋白复合物成分之间的相互作用如何影响其活性和特异性。该提案的具体目标是： 1. 确定 RNase MRP 的蛋白质亚基如何与 RNA 部分相互作用； 2. 确定RNase MRP蛋白组分在酶活性和特异性中的作用。我们将使用 RNA 足迹、突变研究和结构分析来表征 RNase MRP 中的 RNA-蛋白质相互作用，并确定蛋白质部分在 RNase MRP 活性中的作用。这些实验将阐明 RNase MRP 和相关酶中蛋白质的功能。与公共卫生的相关性 RNase MRP 是一种基于 RNA 的酶，是真核细胞生存所必需的。人类 RNase MRP 的 RNA 成分突变会导致软骨毛发发育不全，这是一种严重的多系统疾病，导致侏儒症和免疫缺陷，而对 RNase MRP 的研究有可能揭示该疾病的病因。