Proteins in RNA-Based Enzymes

基于 RNA 的酶中的蛋白质

• 批准号: 7506024
• 负责人: ANDREY S. KRASILNIKOV
• 金额: $ 28.76万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 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/7506024
• 关键词: ABCC1 gene; Affect; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biological Process; Catalysis; Catalytic RNA; Cell Cycle Regulation; Cell Death; Class; Complex; Development; Disease; Dwarfism; Endoribonucleases; 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; Pancreatic ribonuclease; 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; Urination; 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

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学过程中至关重要，包括翻译和RNA处理。尽管在一些基于RNA的酶中，仅RNA部分就能进行催化，但其中许多含有其功能所需的蛋白质。我们的建议集中于核核糖核酸酶MRP（RNase MRP），这是理解蛋白质和RNA在核糖核蛋白蛋白复合物中的相互作用的绝佳模型。 RNase MRP是一种与RRNA成熟和酵母细胞周期的调控有关的真核核糖核蛋白复合物。 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含有十种蛋白质和340个核苷酸长RNA成分。 RNase MRP和RNase P之间的相似性强烈表明RNase MRP中的催化是基于RNA的。但是，与RNase P一样，体内活性也需要所有RNase MRP蛋白。特别有趣的是，这种对RNase P/MRP家族酶中蛋白质的依赖倾向于在更进化的晚期生物体中生长。关于为什么随着进化的进化，RNase P/MRP家族的酶获得越来越多的蛋白质，或者为什么在催化RNA部分似乎已经保留了RNase MRP需要蛋白质的原因。缺乏知识代表了一个问题，因为它阻碍了对这些重要酶的进一步理解。该建议的重点是酿酒酵母核酸链球菌RNase MRP的生化和结构研究，旨在了解该核糖核蛋白复合物的成分之间的相互作用如何影响其活性和特异性。该提案的具体目的是：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蛋白成分在酶的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表征RNA酶MRP中RNA蛋白的相互作用，并确定蛋白部分在RNase MRP活性中的作用。这些实验将阐明蛋白在RNase MRP和相关酶中的功能。与公共卫生RNase MRP相关的是一种基于RNA的酶，这是真核细胞存活所必需的。人类RNase MRP的RNA成分的突变会导致软骨发育不良的发展，严重的多系统疾病导致矮和免疫缺陷，RNase MRP的研究有可能使疾病的原因阐明。