Structure, Function, and Genetics of Ribosomal Components

本项目将重点研究原核核糖体的蛋白质和RNA组分的结构、功能和遗传学。(1)大肠杆菌16S和23S rrna在核糖体组装过程中与r-蛋白S8和L1特异性相互作用的结构特征将通过定点诱变和蛋白质- rrna交联来探测。此外，还将确定蛋白质结合所需的最小rRNA片段，以便将来对其进行详细的物理表征。(2)在大肠杆菌r-蛋白S8无法翻译调控spc操纵子表达的基础上，已分离出40多个突变体。这些蛋白质的代谢和物理稳定性将被检查，并评估其rna结合特性。野生型和突变体S8的二维核磁共振分析将用于高分辨率地定义S8的结构及其rna结合域。(3)对几种嗜热酸性古细菌中翻译和转录装置的编码子进行分离和测序。将采用各种方法来深入了解这些转录单位的表达是如何被控制的。结果将提供一个更好的理解基因调控的进化，以及基因组织，在这一不寻常的生物群体。

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.