Structure, Function, and Genetics of Ribosomal Components

该项目将侧重于结构、功能和 原核生物的蛋白质和RNA成分遗传学 核糖体 (1)E. coli 16 S和 23 S rRNA解释了它们与r- 蛋白质S8和L1在核糖体组装过程中将探测 定点诱变和蛋白质-rRNA交联。 在 此外，蛋白质所需的最小rRNA片段 将确定约束力，以方便其详细说明 未来的物理特性。 (2)超过40个变种人 大肠大肠杆菌R-蛋白S8已被分离的基础上， 它们不能主动调节SPC操纵子 表情这些物质的代谢和物理稳定性 蛋白质将被检查，它们的RNA结合特性 评估。 野生型和突变体S8的2D NMR分析将 用于定义S8的结构及其RNA结合 域，高分辨率。 (3)操纵子编码组件 翻译和转录设备的一些 将分离嗜热嗜酸古细菌， 测序 将采用各种方法来获得 这些转录单位的表达 是被控制的 结果将提供更好的 了解基因调控的进化，以及 基因组织的一个重要组成部分。

This project will focus upon the structure, function and genetics of protein and RNA components of the prokaryotic ribosome. (1) The structural features within E. coli 16S and 23S rRNAs that account for their specific interaction with r- proteins S8 and L1 during ribosome assembly will be probed by site-directed mutagenesis and protein-rRNA cross-linking. In addition, the minimum rRNA segment required for protein binding will be determined to facilitate their detailed physical characterization in the future. (2) Over 40 mutants of E. coli r-protein S8 have been isolated on the basis of their inability to translationally regulate spc operon expression. The metabolic and physically stability of these proteins will be examined, and their RNA-binding properties evaluated. 2D NMR analysis of wild-type and mutants S8 will be used to define the structure of S8, and its RNA-binding domain, at high resolution. (3) Operons encoding components of the translation and transcription apparatus in several thermoacidophilic archaebacteria will be isolated and sequenced. Various approaches will be employed to gain insights into how the expression of these transcription units is controlled. The results will provide a better understanding of the evolution of gene regulation, as well as of gene organization, in this unusual group of organisms.