COORDINATION OF E COLI RIBOSOMAL PROTEIN SYNTHESIS

大肠杆菌核糖体蛋白合成的协调

• 批准号: 2060201
• 负责人: LASSE A LINDAHL
• 金额: $ 18.42万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-07-01 至 1996-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2060201
• 关键词: DNA directed RNA polymerase; Escherichia coli; bacterial genetics; binding proteins; cell growth regulation; gene expression; genetic mapping; genetic promoter element; genetic translation; messenger RNA; molecular cloning; mutant; nucleic acid sequence; operon; protein biosynthesis; ribosomal RNA; ribosomal proteins; scintillation counter; site directed mutagenesis; transcription termination

The synthesis of ribosomes is regulated in response to environment and development. A well-documented example of this is the growth rate control of ribosome synthesis in Escherichia coli. The goal of this project is to unravel the molecular mechanisms underlying this regulation. The information gained by this study should contribute to tbe understanding of regulation of cell growth. It will also elucidate mechanistic aspects of the processes of transcription termination and translation initiation. The E. coli ribosomal protein L-4, encoded by the third gene of the eleven gene S10 operon, is a multifunctional protein which serves as a ribosomal component and as a regulator of both transcription termination in the S10 leader and translation of the first gene of the operon. The purpose of the experiments proposed here is to provide a detailed molecular model for the two regulatory processes induced by excess L4. Currently it is known that transcription termination requires L4, NusA and certain leader sequences, but it is not clear how these components interact to prompt the RNA polymerase to terminate transcription at a specific site. Furthermore, it appears likely that additional component(s) are necessary for completion of the terrmination process. The mechanism for the translation control has not yet been elucidated. A proposed role of alternative secondary RNA structures will be tested and in vitro systems will be established for biochemical analysis of the translation regulation. Finally, L4 mutants will be analyzed to identify domains involved in the protein's various functions. Other experiments will explore new features of the regulation of the S10 operon: (i) possible upstream activating sequences, (ii) possible involvement of sequences outside of the promoter region in steady state growth control, and (iii) expression during stationary phase and out growth.

核糖体的合成是根据环境和 发展。 有充分记录的例子是增长率 控制大肠杆菌中核糖体合成。 目标的目标 项目是要揭示此基础的分子机制 规定。 这项研究获得的信息应有助于 了解细胞生长的调节。 它也将阐明 转录终止过程的机械方面和 翻译启动。 大肠杆菌核糖体蛋白L-4，由第三个基因编码 11个基因S10操纵子是一种多功能蛋白，用作 核糖体成分和作为两个转录终止的调节剂 在S10领导者和操纵子的第一个基因的翻译中。 这 这里提出的实验的目的是提供详细的 多余L4引起的两个调节过程的分子模型。 目前已知转录终止需要L4，NUSA 和某些领导者序列，但尚不清楚这些组件如何 相互作用以提示RNA聚合酶在A处终止转录 特定网站。 此外，似乎有可能 组成部分对于完成地形过程是必需的。 翻译控制的机制尚未阐明。 将测试替代次级RNA结构的拟议作用 并将建立体外系统以进行生化分析 翻译调节。 最后，将分析L4突变体 识别参与蛋白质各种功能的域。 其他实验将探讨S10调节的新功能 操纵子：（i）可能上游激活序列，（ii）可能 稳定状态以外的启动子区域以外的序列参与 生长控制和（iii）在固定阶段和外面表达 生长。