ACTIVATION AND CONTROL OF RRNA TRANSCRIPTION

RRNA 转录的激活和控制

• 批准号: 3070979
• 负责人: Richard L. Gourse
• 金额: $ 5.36万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3070979
• 关键词: DNA binding protein; DNA directed RNA polymerase; DNA footprinting; Escherichia coli; bacterial genetics; genetic promoter element; genetic transcription; mutant; nucleic acid sequence; protein biosynthesis; ribosomal RNA; ribosomes

In E. coli, the synthesis of ribosomes is controlled primarily at the stage of rRNA transcription initiation. rRNA can account for as much as 50% of instantaneous cellular RNA synthesis. In order to understand the biosynthesis of the protein synthetic apparatus, factors involved in the upstream activation and growth rate dependent regulation of rRNA synthesis are being investigated. The mechanism of rRNA promoter-RNA polymerase association is being studied using kinetic and footprinting methods. This will lead to the identification of the individual steps involved in transcription initiation at the rrnB PL promoter and the factors that influence each step. The specific DNA sequences required for upstream activation and growth rate control will be defined by construction and selection of site mutations, followed by assay of operon fusions in vivo. Finally, the importance of position, orientation, and conformation of the upstream activator sequence (UAS) relative to the promoter and the feasibility of using the UAS to enhance transcription from heterologous promoters will also be examined using operon fusions in vivo.

在大肠在大肠杆菌中，核糖体的合成主要控制在 rRNA转录起始阶段。 rRNA可以解释为 高达50%的瞬时细胞RNA合成。 为了 了解蛋白质合成装置的生物合成， 上游激活和生长速率的相关因素 rRNA合成的依赖性调节正在研究中。 rRNA启动子-RNA聚合酶结合的机制是 正在使用动力学和足迹法进行研究。 这将 导致识别所涉及的各个步骤， 转录起始于rrnB PL启动子， 影响每一步。 所需的特定DNA序列 对于上游激活和生长速率控制， 构建和选择位点突变，然后测定 体内操纵子融合。 最后，位置的重要性， 上游激活序列的方向和构象 (UAS)相对于启动子和使用的可行性 UAS增强异源启动子的转录将 也可以在体内使用操纵子融合进行检查。