MECHANISM & ACTIVATION OF RRNA TRANSCRIPTION

机制

• 批准号: 3291946
• 负责人: Richard L. Gourse
• 金额: $ 14.27万
• 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: 1988-07-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3291946
• 关键词: DNA directed RNA polymerase; DNA footprinting; Escherichia coli; bacterial genetics; genetic manipulation; 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 asociation 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 P1 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，核糖体的合成主要在 rRNA转录起始的关键。 rRNA可以占到50%， 瞬时细胞RNA合成。 为了解 蛋白质合成装置的生物合成，参与蛋白质合成的因素， rRNA合成的上游激活和生长速率依赖性调节 正在接受调查 rRNA启动子-RNa聚合酶的作用机制 使用动力学和足迹法研究关联。 这 将导致识别所涉及的各个步骤， 转录起始于rrnB P1启动子， 影响每一步。 上游所需的特定DNA序列 激活和增长率控制将由施工和 位点突变的选择，随后进行操纵子体内融合的测定。 最后，阐述了企业文化的定位、定位和形态的重要性， 上游激活子序列（UAS）， 使用UAS增强异源转录的可行性 还将使用操纵子融合体在体内检查启动子。