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

这里提出的研究涉及共价连接Fe（II）-EDTA 核糖体RNA（rRNA）探针，以确定三级 RNA的高度保守区域的结构。 它是一个更大的 该项目旨在制作最精确的三维模型， 大肠杆菌核糖体。 核糖体进行蛋白质合成， 生活中最基本的方面之一。 除其他功能外，蛋白质 合成已被证明是细胞周期的一部分。 知道 核糖体的三级结构将有助于阐明 核糖体和翻译机制，这应该有助于发现 阻止细胞周期并提供癌症治疗的方法。 两 本文提出了Fe（Ⅱ）-EDTA与rRNA结合的方法：1）Fe（Ⅱ）-EDTA 将被连接到5 '-硫代磷酸酯，从而在 核糖核酸 2)Fe（II）-EDTA将连接到修饰的尿苷上， 一个连续的RNA分子 修饰RNA的切割模式 将用于确定23 S rRNA的哪些部分与16 S rRNA接触 在核糖体的30 S和50 S亚基之间的界面处， 确定16 S rRNA的哪些区域靠近530茎环， 具体地确定1400-1500解码区域是否接近于 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.