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（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片段能够单独发挥催化作用，但其中许多酶含有实现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)，这是一个很好的模型来理解蛋白质和RNA在核糖核蛋白复合物中的相互作用。RNase MRP是一种真核核糖核蛋白复合物，在酵母中参与rRNA的成熟和细胞周期的调节。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包含10个蛋白和340个核苷酸长的RNA组分。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的情况一样，所有RNase MRP蛋白也需要在体内具有活性。特别有趣的是，这种对RNase P/MRP家族酶中的蛋白质的依赖倾向于在更进化的高级生物体中生长。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RNase MRP需要蛋白质，而催化RNA片段似乎基本上被保留了下来，这一点我们知之甚少。这种知识的缺乏代表了一个问题，因为它阻碍了对这些重要酶的进一步了解。本课题主要对酿酒酵母核核RNase MRP进行生化和结构研究，旨在了解该核糖核蛋白复合物组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片段相互作用；2. 确定RNase MRP蛋白组分在酶的活性和特异性中的作用。我们将利用RNA足迹、突变研究和结构分析来表征RNase MRP中RNA-蛋白质的相互作用，并确定蛋白质片段在RNase MRP活性中的作用。这些实验将阐明这些蛋白在RNase MRP及相关酶中的功能。rna酶MRP是一种真核细胞生存所必需的rna酶。人类RNase MRP的RNA成分突变导致软骨毛发育不全，这是一种严重的多系统疾病，导致侏儒症和免疫缺陷，对RNase MRP的研究有可能揭示这种疾病的原因。