CAREER: Defining the Chemical Basis of RNA Function by Nucleotide Analog Interference Mapping

Strobel教授的研究重点是建立一种新的方法， 在结构上确定构效关系， 催化重要的RNA分子。 靶分子包括一系列 具有氧化不稳定的5 '-O-（1-硫代）三磷酸的核苷酸类似物 取代基。 单个类似物将通过插入被并入RNA中。 体外转录以创建突变RNA文库，其中个体 核苷酸被类似物随机替换。 个别RNA 在特定生物系统中表现出活性的突变体是 区别于那些没有的。 随后的氧化裂解， 在温和条件下与碘反应， 活性和非活性类似物在测序上的比较分析 凝胶提供了一种直接的方法来识别活性化学物质 组 这种方法，被称为核苷酸抗干扰， NAIM）将用于识别RNA中的化学基团 涉及：（1）5SrRNA与核糖体蛋白L25的相互作用; (2)U6 snRNA在前体mRNA剪接中的作用;（3）自催化VS 脉孢菌属的核酶;（4）α操纵子mRNA的相互作用 S4蛋白 教育活动包括发展一个 生物有机课程强调合成有机设计和大分子 功能，并涉及当地本科生和少数民族学生 在研究项目中。 有了这个职业奖，有机动力学计划支持 研究和教育活动的教授斯科特斯特罗贝尔的 耶鲁大学分子生物物理与生物化学系。 斯特罗贝尔教授的研究重点是开发一种方法， 称为核苷酸标记干涉图，这将显着 加快确定结构-活性关系的进程， 结构和催化重要的RNA分子。 教授 斯特罗贝尔的教育活动包括开发一个多部门的 课程将探讨有机化学合成的使用， 研究和理解生物系统，并促进兴趣 在化学研究中，当地和少数民族大学生都参与其中， 在研究项目中。

Professor Strobel's research focuses on establishing a new methodology to determine structure-activity relationships in structurally and catalytically important RNA molecules. Target molecules include a series of nucleotide analogues with oxidatively labile 5'-O-(1-thio)triphosphate substituents. Individual analogues will be incorporated into RNA by in vitro transcription to create a library of mutant RNA's, where individual nucleotides have been randomly replaced by an analogue. Individual RNA mutants that exhibit activity in a particular biological system are distinguished from those that do not. Subsequent oxidative cleavage at the thiophosphate group under mild conditions with iodine, and comparative analysis of the active and nonactive analogues on sequencing gels offers a straightforward means to identify the active chemical group. This methodology, called Nucleotide Analogue Interference Mapping (NAIM), will be used to identify the chemical groups in RNA relating to: (1) the interaction of 5S rRNA with ribosomal protein L25; (2) the role of U6 snRNA in pre-mRNA splicing; (3) the autocatalytic VS ribozyme from Neurospora; and (4) the interaction of alpha operon mRNA with S4 protein. Educational activities include developing a bioorganic course emphasizing synthetic organic design and macromolecular function, and involving both local undergraduates and minority students in the research program. With this CAREER award, the Organic Dynamics Program supports the research and educational activities of Professor Scott Strobel of the Department of Molecular Biophysics and Biochemistry at Yale University. Professor Strobel's research is focused on developing a methodology, called Nucleotide Analogue Interference Mapping, that will significantly accelerate the process of identifying structure-activity relationships in structurally and catalytically important RNA molecules. Professor Strobel's educational activities include developing a multidepartmental course which will explore the use of organic chemical synthesis to investigate and understand biological systems, and promoting an interest in chemical research by involving both local and minority undergraduates in the research program.