RNA REGULATORY DOMAINS: FOLDING, RECOGNITION & CATALYSIS

RNA 调控域：折叠、识别

• 批准号: 7001236
• 负责人: DINSHAW J PATEL
• 金额: $ 32.7万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7001236
• 关键词: X ray crystallography; adenine; allosteric site; bioenergetics; calorimetry; catalyst; cell cycle; chemical binding; chemical cleavage; chemical structure function; gene expression; gene mutation; guanine; histones; intermolecular interaction; messenger RNA; nuclear magnetic resonance spectroscopy; nucleic acid sequence; protein degradation; ribozymes; surface plasmon resonance; thiamine pyrophosphate

DESCRIPTION (provided by applicant): This revised application focuses on 3 projects addressing structural and functional aspects of folding, recognition and catalysis by RNA regulatory domains. Project 1 focuses on the structural characterization of the core domains of metabolite-sensing mRNAs discovered in the Ronald Breaker laboratory, which adopt complex junctional topologies capable of ligand-induced functional modulation. Our structural research will initially focus on the free and bound core domains of guanine/adenine-sensing and thiamine pyrophosphate-sensing mRNAs, for which we have collected promising NMR and crystallographic data. Our structural studies combined with functional efforts in the Breaker laboratory should highlight the principles of molecular recognition and metabolite encapsulation by mRNA, and define the allosteric mRNA transitions associated with the modulation of gene expression levels and metabolic homeostasis. Project 2 on ribozymes catalyzing chemical reactions, focuses on the structural characterization of RNA motifs with Diels-Alderase catalytic activities discovered in the Andres Jaschke laboratory. We are attempting to structurally characterize the catalytic RNA scaffold in the context of bound substrates, transition state analogs and products, with promising crystallographic and NMR data collected on the product complex. Our structural characterization of the Diels-Alderase ribozyme, together with mutational and energetics studies in the Jaschke laboratory, should identify principles for generation of novel ribozymes with controllable catalytic activities, tunable specificites and enantiomeric capabilities. Project 3 focuses on protein-RNA recognition events that mediate the degradation of metazoan histone mRNA, a process tightly coupled to cell cycle progression. Histone mRNAs contain a unique bipartite stem-loop scaffold followed by an ACCA sequence, whose stem, loop and flanking sequences are targeted along opposite faces, both separately and simutaenously, by a stem-loop binding protein and a histone mRNA 3' end-specific exonuclease. Our proposed structural and energetics characterization of binary and ternary protein-RNA recognition events involved in histone mRNA 3'-end recognition and cleavage should provide insights into the mechanisms that cells use to achieve precise cell cycle-regulated mRNA degradation.

描述（由申请人提供）：此修订后的申请重点是3个项目，这些项目涉及RNA调节域的折叠，识别和催化的结构和功能方面。项目1的重点是在Ronald Breaker实验室中发现的代谢产物感应mRNA的核心域的结构表征，该实验室采用了能够配体诱导的功能调制的复杂连接拓扑。我们的结构研究最初将集中于鸟嘌呤/腺嘌呤感应和硫胺素焦磷酸含量的mRNA的自由和界限核心结构域，为此，我们收集了有希望的NMR和晶体学数据。我们的结构研究结合了断路器实验室中的功能努力，应强调mRNA分子识别和代谢物封装的原理，并定义与基因表达水平和代谢稳态的调节相关的变构mRNA转变。核酶催化化学反应的项目2，重点介绍了在安德烈斯·贾斯克（Andres Jaschke）实验室中发现的Diels-碱酶催化活性的RNA基序的结构表征。我们正在尝试在结构底物，过渡状态类似物和产物的背景下在结构上表征催化RNA支架，并在产品复合物上收集了有希望的晶体学和NMR数据。我们对DIELS-积极酶核酶的结构表征，以及Jaschke实验室中的突变和能量研究，应确定具有具有可控催化活性，可调的特定材料和对映体功能的新型核酶的原理。项目3的重点是蛋白-RNA识别事件，这些事件介导了后生组组蛋白mRNA的降解，这一过程与细胞周期的进程紧密耦合。组蛋白mRNA包含一个独特的二分线杆脚架，然后是ACCA序列，其茎，环和侧翼序列沿对面的面孔分别和同样是通过茎环结合蛋白和组蛋白mRNA 3'末端特异性外核酸酶的靶向。我们提出的针对组蛋白mRNA 3'-End识别和裂解的二元和三元蛋白RNA识别事件的结构和能量表征应提供对细胞用于实现精确细胞周期受到的mRNA降解的机制的见解。