Structure of ribonucleoprotein: telomerase

核糖核蛋白的结构：端粒酶

• 批准号: 6777715
• 负责人: Gabriele Varani
• 金额: $ 29.74万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-09 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6777715
• 关键词: RNA; RNA directed DNA polymerase; X ray crystallography; conformation; enzyme activity; enzyme complex; enzyme mechanism; enzyme structure; fungal proteins; intermolecular interaction; molecular assembly /self assembly; nuclear magnetic resonance spectroscopy; posttranscriptional RNA processing; protein protein interaction; protein purification; protein quantitation /detection; protein structure function; ribonucleoproteins; telomerase; telomere; yeasts

DESCRIPTION (provided by applicant): The goal of this proposal is to study the structure/function relationship of telomerase. This enzyme protects the ends of chromosomes, maintains their length during cellular division, and is selectively activated in most humans cancers. Through these activities, it maintains genomic integrity, promotes cellular immortalization, and sustains the proliferating status of transformed cells: its pharmacological inhibition may very well provide a new avenue to cancer treatment. Telomerase is a ribonucleoprotein composed of a non-coding RNA, a protein enzymatic component and additional essential protein subunits. Very little is known structurally on telomerase or its components, yet this knowledge, and an understanding of the molecular interactions between its protein and RNA components, are essential to understand its function and mechanism. We propose to determine the structure of telomerase-associated proteins and of key functional domains of its RNA component. The structural information will be integrated into a biochemical and functional context, through collaborations with biochemists and molecular biologists (Gottchling, Lingner and Rhodes), who will provide the breadth of expertise required to answer, at the molecular level, fundamental questions on how this enzyme functions. The enzymatic activity of telomerase (TERT) is homologous to viral reverse transcriptases, while the RNA (TER) provides the template and a landing dock for RNP assembly. Different biochemical functions map to specific domains of TER and to the proteins that associate with them. Because the telomerase holoenzyme is large and complex, I propose to build up its structure from its constituent parts, based on the "divide-and-conquer" principle. Specifically, I propose to study protein-RNA complexes responsible for: 1. RNA processing, stability and assembly of the mature RNP; 2. Targeting of the mature telomerase to the telomeres; 3. Binding and activation of TERT. The research plan will be based on: 1. The analysis by NMR of the structure and dynamics of the domains of TER responsible for each of these functions; 2. The determination of the structure of the proteins that interact with such domains; 3. The biochemical and structural characterization of the resulting RNA-protein complexes; 4. The design and execution of new biochemical and functional experiments, grounded onto the structural results, that will address the biological function of these protein-RNA complexes.

描述（由申请人提供）：本提案的目标是研究端粒酶的结构/功能关系。这种酶可以保护染色体末端，在细胞分裂过程中保持其长度，并且在大多数人类癌症中被选择性激活。通过这些活动，它维持基因组完整性，促进细胞永生化，并维持转化细胞的增殖状态：其药理学抑制很可能为癌症治疗提供新途径。端粒酶是一种核糖核蛋白，由非编码 RNA、蛋白质酶促成分和其他必需蛋白质亚基组成。关于端粒酶或其组分的结构知之甚少，但这些知识以及对其蛋白质和 RNA 组分之间分子相互作用的理解对于理解其功能和机制至关重要。我们建议确定端粒酶相关蛋白及其 RNA 成分关键功能域的结构。 通过与生物化学家和分子生物学家（Gottchling、Lingner 和 Rhodes）的合作，结构信息将被整合到生化和功能背景中，他们将提供所需的广泛专业知识，以在分子水平上回答有关这种酶如何发挥作用的基本问题。功能。 端粒酶（TERT）的酶活性与病毒逆转录酶同源，而RNA（TER）则为RNP组装提供模板和着陆点。不同的生化功能映射到 TER 的特定域以及与其相关的蛋白质。由于端粒酶全酶庞大且复杂，我建议根据“分而治之”的原则，从其组成部分构建其结构。具体来说，我建议研究蛋白质-RNA复合物，其负责： 1. RNA加工、成熟RNP的稳定性和组装； 2. 将成熟的端粒酶靶向端粒； 3.TERT的结合和激活。研究计划将基于： 1. 通过 NMR 对负责这些功能的 TER 域的结构和动力学进行分析； 2. 确定与这些结构域相互作用的蛋白质的结构； 3. 所得RNA-蛋白质复合物的生化和结构表征； 4. 以结构结果为基础，设计和执行新的生化和功能实验，以解决这些蛋白质-RNA 复合物的生物学功能。