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排列的技术进行研究， 它与mRNA、tRNA和核糖体蛋白的相互作用。 本研究将 导致了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 互补技术来研究rRNA的功能。 这些 实验将为理解mRNA是如何 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为定点光亲和剂。 因为我们可以控制 所测量的距离的放置将大约为5A。 UV 交联也将被用来监测的整体配置， 16S rRNA。 这两种技术对结构非常敏感。 变化 这项工作的具体目标是： 1. 为了确定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. 为了确定亚基结合、mRNA结合和tRNA 结合诱导16S rRNA的三维结构变化， 3. 为了确定野生型结构如何被16S突变改变， 影响mRNA和tRNA结合的rRNA。 4. 为了确定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.