An Unbiased Chemical Approach for the Discovery of Naturally Occurring Ribozymes

发现天然核酶的公正化学方法

• 批准号: 8536647
• 负责人: Richard Ian McDonald
• 金额: $ 5.22万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-03 至 2014-08-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8536647
• 关键词: Affinity; Amides; Binding; Biochemical; Biological; Biological Process; Biotin; Catalysis; Catalytic RNA; Cell physiology; Cells; Chemicals; Collection; Defense Mechanisms; Detection; Development; Disease; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Gene Expression Regulation; Genetic; Genome; Human; Hydrolysis; In Vitro; Investigation; Label; Lead; Link; Methodology; Methods; Organism; Pathway interactions; Play; Proteins; RNA; RNA Processing; Relative (related person); Research; Ribosomes; Role; Structure; Untranslated RNA; activity-based protein profiling; aptamer; base; catalyst; chemical reaction; design; functional group; genome-wide; novel; phosphodiester; screening; small molecule; tool

DESCRIPTION (provided by applicant): The discovery that RNA can catalyze chemical reactions revealed a novel role for these molecules that extends beyond the transfer of genetic information. To date, only a small number of biologically occurring catalytic RNAs (ribozymes) have been identified. This may be a result of the absence of unbiased, activity- based methods for their identification. Approaches that do not rely on secondary structure prediction and instead select RNAs possessing enhanced reactivity could reveal novel ribozymes. This proposal describes the development of a chemical approach to the discovery of biological ribozymes. This activity-based approach employs small-molecule probes designed to covalently label RNAs possessing enhanced chemical reactivity. The strategy for the design of ribozyme-specific chemical probes relies on the well-validated assumption that ribozymes employ similar tools as protein catalysts; the tertiary structures of most ribozymes result in functional groups with enhanced nucleophilicity that are largely responsible for catalysis. Appropriately designed electrophilic compounds (activity-based probes) designed to selectively label highly nucleophilic RNAs will be equipped with an affinity tag (biotin) and applied to the selection of unusually reactive RNAs from RNA pools derived from the genomes of a range of bacterial and eukaryotic organisms. The chemical reactivity and biological roles of any newly identified RNAs will be fully characterized.

描述（由申请人提供）：发现RNA可以催化化学反应的发现揭示了这些分子的新作用，该作用超出了遗传信息的转移。迄今为止，仅鉴定出少量的生物学发生的催化RNA（核酶）。这可能是由于缺乏公正的基于活动的方法来识别其。不依赖二级结构预测的方法，而是 具有增强反应性的选择RNA可以揭示新的核酶。 该提案描述了一种化学方法来发现生物核酶。这种基于活性的方法采用了旨在共价标记具有增强化学反应性的RNA的小分子探针。核酶特异性化学探针设计的策略依赖于验证核酶采用与蛋白质催化剂相似的工具的验证假设。大多数核酶的三级结构导致具有增强的亲核性的官能团，这在很大程度上是导致催化的原因。适当设计的亲电化合物（基于活性的探针），设计用于选择性标记高度亲核RNA的高度核RNA将配备亲和力TAG（Biotin），并应用于从来自细菌和真核生物生物体的一系列基因组中得出的异常反应性RNA的选择。任何新确定的RNA的化学反应性和生物学作用都将得到充分表征。