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 用确定rRNA和rRNA排列的技术进行研究 它与mRNA、tRNA和核糖体蛋白的相互作用。这项研究将 以更高的分辨率获得核糖体的三维模型 目前是可用的。定点突变将被用作一种 研究rRNA功能的补充技术。这些 实验将为理解信使核糖核酸是如何 选择，如何确定蛋白质合成的速度和终止 以及抗生素是如何破坏核糖体功能的。 结构分析将主要用特定部位的补骨脂素进行 (SSP)，k是一种定点光亲和剂。因为我们可以控制 所测量的距离的放置将大约为5A。 交叉链接还将用于监控的整体配置 16S rRNA。这两种技术都将对结构非常敏感 改变。 这项工作的具体目标是： 1.确定16S rRNA的三维结构。 活化的30S亚基和70S核糖体中的mRNA解码位点， 2.确定亚单位结合、mrna结合和trna 结合诱导16S rRNA的三维结构变化， 3.确定16s突变对野生型结构的影响 影响mRNA和tRNA结合的rRNA。 4.确定核糖体中信使核糖核酸的途径。 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.