Proteins in RNA-Based Enzymes

基于 RNA 的酶中的蛋白质

• 批准号: 8269726
• 负责人: ANDREY S. KRASILNIKOV
• 金额: $ 28.29万
• 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 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8269726
• 关键词: Affect; Binding Sites; Biochemical; 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; Structural Biochemistry; Therapeutic Agents; Time; Transfer RNA; Translation Process; Translations; X-Ray Crystallography; Yeasts; base; cartilage development; enzyme activity; human disease; in vivo; mRNA Transcript Degradation; novel therapeutics; 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(申请人提供)：基于RNA的酶在许多重要的生物过程中都是至关重要的，包括翻译和RNA加工。虽然在一些基于RNA的酶中，只有RNA部分能够执行催化作用，但许多酶含有发挥其功能所需的蛋白质。我们的建议侧重于核核糖核酸酶MRP(RNase MRP)，这是理解核糖核蛋白复合体中蛋白质和RNA相互作用的一个很好的模型。核糖核酸酶MRP是一种真核核糖核蛋白复合体，参与酵母中rRNA的成熟和细胞周期的调节。核糖核酸酶MRP与核糖核酸酶P关系密切，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个核苷酸的长RNA组分。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；然而，与RNase P一样，所有RNase MRP蛋白也是体内活性所必需的。特别耐人寻味的是，这种对RNaseP/MRP家族酶中蛋白质的依赖往往会在更进化的高级生物体中生长。人们对RNase P/MRP家族的酶为什么随着进化获得越来越多的蛋白质，或者为什么RNase MRP需要蛋白质，而催化的RNA部分似乎基本上被保存下来，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。本建议重点研究酿酒酵母核仁RNase MRP的生化和结构，旨在了解这种核糖核蛋白复合体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的蛋白质亚单位如何与RNA部分相互作用；2.确定RNase MRP蛋白质组分在酶的活性和特异性中的作用。我们将通过RNA足迹、突变研究和结构分析来表征RNase MRP中RNA与蛋白质的相互作用，并确定蛋白质部分在RNase MRP活性中的作用。这些实验将阐明这些蛋白质在RNase、MRP和相关酶中的功能。与公共卫生相关核糖核酸酶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成分的突变会导致软骨头发发育不良，这是一种严重的多系统疾病，导致侏儒症和免疫缺陷，对RNase MRP的研究有可能揭示这种疾病的原因。