Transcription termination and its regulation in E. Coli

大肠杆菌中的转录终止及其调控

基本信息

批准号：
8672454
负责人：
EVGENY A NUDLER
金额：
$ 16.95万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8672454
关键词：
Address Amino Acids Antibiotics Bacterial Genes Bacterial RNA Bacteriophages Binding Sites Biochemical Carbon Cell Survival Cells ChIP-on-chip Chemicals Complex Consensus Coupled DNA DNA-Directed RNA Polymerase Elongation Factor Enterobacter Escherichia coli Essential Genes Future Gene Expression Gene Expression Regulation Genes Genetic Transcription Genomics Goals Grant Growth Health Human Intrinsic factor Kinetics Maps Microarray Analysis Modeling Molecular Mutation Nutrient Pathway interactions Pattern Pharmaceutical Preparations Phenotype Physiological Process Protein Inhibition Proteins RNA RNA Binding Regulation Research Resistance Rho Factor Role Signal Transduction Site Source Structure Testing Thermodynamics Transcript Translations Vitamins Work antimicrobial antitermination bicozamycin chromatin immunoprecipitation design dietary supplements environmental change genome-wide in vivo inhibitor/antagonist insight multidisciplinary novel protein protein interaction research study response rho small molecule termination factor transcription termination

项目摘要

DESCRIPTION (provided by applicant): Transcription termination is a process whereby the elongation complex (EC) dissociates into RNA transcript, DNA template, and RNA polymerase (RNAP) in response to intrinsic signals or specific factors. Rho termination factor is essential in regulating gene expression in Enterobacter and is a target for specific antibiotics. Rho has been intensively studied in the last three decades, however, the actual termination process, i.e. the mechanism by which Rho disrupts the EC, remains unknown. Moreover, the identity of most Rho termination sites in vivo and the role of Rho in cell's adaptation to environmental changes remain unknown. The long-term objective of the proposed work is to provide a comprehensive physiological and mechanistic description of Rho-dependent termination in Escherichia coli and the mechanism of its regulation by particular host and phage proteins. Specifically we propose to: 1) Determine conformational changes in RNAP that accompany the termination process, and the role of certain RNAP domains in Rho termination. 2) Determine how E.coli S4 and phage l N proteins modify the EC rendering it resistant to Rho termination. 3) Determine the physiological role and mechanism of novel anti-Rho factors that we have identified in preliminary studies. 4) Establish the role of Rho in gene regulation on a genomic scale. The significance of proposed research for human health is several-fold. Complete structural understanding of termination/antitermination processes would allow for designing small molecule mimics and inhibitors that change the pattern of bacterial gene expression or interrupt transcription of essential genes prematurely, and thus serve as novel antimicrobials. Examples of antibiotics that specifically target Rho have been already described. Furthermore, better understanding the mechanisms of antitermination would suggest the optimal strategies for constructing bacterial strains that overproduce essential dietary supplements and other biologically active compounds. Finally, since eukaryotic RNAPs share basic sequence and structural homologies with bacterial RNAP, the fundamental mechanism of the EC stabilization and destabilization must be similar. Therefore, proposed experiments will also provide insight to the basic mechanisms of eukaryotic transcription termination. We will explore the mechanism of Rho termination and antitermination in E.coli. The significance of proposed research for human health is several-fold. Complete structural understanding of termination/antitermination processes would allow designing small molecule mimics and inhibitors of the termination process that change the pattern of bacterial gene expression or interrupt transcription of essential genes prematurely, and thus serve as novel antimicrobials. Furthermore, better understanding the mechanisms of antitermination would suggest optimal strategies for constructing bacterial strains that overproduce essential dietary supplements (vitamins, amino acids, etc.) and other important biologically active compounds. Finally, the proposed experiments will also provide insight to the basic mechanisms of eukaryotic transcription termination.

描述（由申请人提供）：转录终止是一个过程，延伸复合物（EC）将响应内在信号或特定因素响应RNA转录物，DNA模板和RNA聚合酶（RNAP）中。 RHO终止因子对于调节肠杆菌中的基因表达至关重要，并且是特定抗生素的靶标。在过去的三十年中，对Rho进行了深入研究，但是，实际终止过程，即Rho破坏EC的机制仍然未知。此外，大多数RHO终止位点在体内的身份以及Rho在细胞适应环境变化中的作用尚不清楚。拟议工作的长期目标是提供大肠杆菌中Rho依赖性终止的全面生理和机械描述，以及其对特定宿主和噬菌体蛋白的调节机制。具体而言，我们建议：1）确定终止过程中RNAP的构象变化，以及某些RNAP域在RHO终止中的作用。 2）确定大肠杆菌S4和噬菌体l n蛋白如何改变其对Rho终止的抗性。 3）确定我们在初步研究中确定的新型抗RHO因素的生理作用和机制。 4）在基因组量表上确定RHO在基因调节中的作用。拟议的人类健康研究的意义是多个。对终止/抗释放过程的完整结构理解将允许设计小分子模拟物和抑制剂，以改变细菌基因表达的模式或过早的必需基因的中断转录，从而作为新型抗菌剂。已经描述了专门针对RHO的抗生素的例子。此外，更好地理解抗授权的机制将提出构造细菌菌株的最佳策略，从而过量生产必需的饮食补充剂和其他生物学活性化合物。最后，由于真核RNAP具有细菌RNAP的基本序列和结构同源性，因此EC稳定和不稳定的基本机制必须相似。因此，提出的实验还将为真核转录终止的基本机制提供见解。我们将探讨大肠杆菌中RHO终止和抗验证的机制。拟议的人类健康研究的意义是多个。对终止/抗释放过程的完整结构理解将允许设计小分子模拟物和抑制剂的终止过程，这些过程会改变细菌基因表达的模式或过早的必需基因的中断转录，从而用作新型抗菌素。此外，更好地理解抗授权的机制将提出最佳策略，以构建过量产生必需饮食补充剂（维生素，氨基酸等）的细菌菌株和其他重要的生物活性化合物。最后，提出的实验还将为真核转录终止的基本机制提供见解。