TETHERED FE(II)--A STRUCTURAL PROBE OF RIBOSOMAL RNA

本文提出的研究涉及共价连接Fe(II)-EDTA 以核糖体RNA(RRNA)为探针，以确定第三代 RNA高度保守区域的结构。它是一个更大的 旨在制作最精确的三维模型的项目 到目前为止，大肠杆菌核糖体。核糖体执行蛋白质合成，这是 生活中最基本的方面之一。在其他功能中，蛋白质 合成已被证明是细胞周期的一部分。了解 核糖体的三级结构将有助于阐明其活性部位 核糖体和翻译机制，这应该有助于发现 阻止细胞周期并为癌症提供治疗的方法。二 提出了将Fe(II)-EDTA固定在rRNA上的方法：1)Fe(II)-EDTA 将附着在5‘-硫代磷酸酯上，从而在 核糖核酸。2)将Fe(II)-EDTA连接到修饰的尿苷生成物上 一种连续的核糖核酸分子。修饰后的RNA的切割模式 将用于确定23S rRNA的哪些部分与16S rRNA接触 在核糖体的30s和50s亚基之间的界面上， 确定16S rRNA的哪些区域靠近530茎环， 具体确定1400-1500编码区是否在 这个循环。

The research proposed here involves covalently attaching Fe(II)-EDTA probes to ribosomal RNA (rRNA) in order to determine the tertiary structure of highly conserved regions of the RNA. It is part of a larger project designed to produce the most accurate three dimensional model of E.Coli ribosomes to date. Ribosomes perform protein synthesis, which is one of the most basic aspects of life. Among other functions, protein synthesis has been shown to be part of the cell cycle. Knowing the tertiary structure of ribosomes will help elucidate the active sites of ribosomes and the mechanism of translation, which should aid in finding ways to arrest the cell cycle and provide a treatment for cancer. Two methods of attaching Fe(II)-EDTA to rRNA are proposed: 1) Fe(II)-EDTA will be attached to a 5'-thiophosphate creating a discontinuity in the RNA. 2) Fe(II)-EDTA will be attached to a modified uridine generating a continuous RNA molecule. The cleavage patterns of the modified RNA will be used to determine what parts of the 23S rRNA contact the 16S rRNA at the interface between the 30S and 50S subunits of ribosomes, and to ascertain what regions of 16S rRNA are close to the 530 stem-loop, specifically determining if the 1400-1500 decoding region is proximal to this loop.