Structure, Function, and Genetics of Ribosomal Components

核糖体成分的结构、功能和遗传学

• 批准号: 9108104
• 负责人: Robert Zimmermann
• 金额: $ 25.5万
• 依托单位: University of Massachusetts Amherst
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9108104&HistoricalAwards=false
• 关键词: Structure; Function; Genetics; Ribosomal; Components

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.

该项目将重点研究原核核糖体蛋白质和 RNA 成分的结构、功能和遗传学。 (1) 将通过定点诱变和蛋白质-rRNA 交联来探测大肠杆菌 16S 和 23S rRNA 内的结构特征，这些结构特征解释了它们在核糖体组装过程中与 r 蛋白 S8 和 L1 的特异性相互作用。 此外，还将确定蛋白质结合所需的最小 rRNA 片段，以方便将来对其进行详细的物理表征。 (2) 超过 40 个大肠杆菌 r 蛋白 S8 突变体已被分离出来，因为它们不能翻译调节 spc 操纵子的表达。将检查这些蛋白质的代谢和物理稳定性，并评估它们的 RNA 结合特性。 野生型和突变体 S8 的 2D NMR 分析将用于以高分辨率定义 S8 的结构及其 RNA 结合域。 (3) 将分离编码几种嗜热嗜酸古细菌中翻译和转录装置成分的操纵子并进行测序。 将采用各种方法来深入了解如何控制这些转录单元的表达。 这些结果将有助于更好地理解这一不寻常生物体中基因调控以及基因组织的进化。