STRUCTURE AND MECHANISM OF RIBOSOMAL RNA IN THE RIBOSOME

核糖体的结构和蛋白质合成的机制将 可以通过技术来研究确定 rRNA 的排列 它与 mRNA、tRNA 和核糖体蛋白的相互作用。 这项研究将 产生更高分辨率的核糖体三维模型 目前可用。 定点诱变将被用作 互补技术研究 rRNA 的功能。 这些 实验将为理解 mRNA 的运作方式提供更好的基础 选择，如何确定蛋白质合成的速率和终止 以及抗生素如何破坏核糖体功能。 结构分析将主要使用位点特异性补骨脂素进行 （SSP），k 一种定点光亲和剂。 因为我们可以控制 测量的距离的放置大约为 5 A。 UV 交叉链接也将用于监控整体配置 16S rRNA。 这两种技术都对结构非常敏感 变化。 这项工作的具体目标是： 1. 确定16S rRNA的三维结构 活性 30S 亚基和 70S 核糖体中的 mRNA 解码位点， 2. 确定是否有亚基关联、mRNA结合和tRNA 结合诱导 16S rRNA 的三维结构变化， 3.确定16S突变如何改变野生型结构 rRNA 影响 mRNA 和 tRNA 结合。 4. 确定mRNA通过核糖体的路径 翻译。

The structure of the ribosome and the mechanism of protein synthesis will be studied with techniques the determine the arrangement of the rRNA and its interaction with mRNA, tRNA, and ribosomal proteins. This study will lead to three dimensional models of the ribosome at higher resolution that is presently available. Site directed mutagenesis will be used as a complementary technique to investigate the function of the rRNA. These experiments will provide a better basis for understanding how mRNA is selected, how the rate and termination of protein synthesis are determined and how antibiotics disrupt ribosome function. The structural analysis will be done primarily with site specific psoralen (SSP),k a site-directed photoaffinity agents. Because we can control the placement of the distances that are measured will be approximately 5 A. UV cross-linking will also be used to monitor the overall configuration of the 16S rRNA. Both of these techniques will be very sensitive to structural changes. The specific aims of this work are: 1. to determine the three dimensional structure of the 16S rRNA at the mRNA decoding site in active 30S subunits and in 70S ribosomes, 2. to determine whether subunit association, mRNA binding, and tRNA binding induce three dimensional structural changes in he 16S rRNA, 3. to determine how the wild type structure is altered by mutations in 16S rRNA that affect mRNA and tRNA binding. 4. to determine the path of the mRNA through the ribosome during translation.