Phage-induced modifications of RNA polymerase

噬菌体诱导的 RNA 聚合酶修饰

• 批准号: 7933443
• 负责人: KONSTANTIN V SEVERINOV
• 金额: $ 25.75万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933443
• 关键词: ADP ribosylation; Affect; Asia; Bacteria; Bacterial RNA; Bacteriophage T4; Bacteriophage T7; Bacteriophage lambda; Bacteriophages; Biological; Biological Assay; Biological Models; Biology; DNA; DNA Packaging; DNA-Directed RNA Polymerase; Development; Drug Delivery Systems; Drug Design; Escherichia coli; Gene Expression; Gene Expression Regulation; Genetic; Genetic Transcription; Genomics; Goals; Hand; In Vitro; Lead; Modification; Molecular; Molecular Probes; Organism; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Proteins; Regulation; Research; Research Personnel; Role; Salmonella typhimurium; Site; Staging; System; Transcriptional Regulation; Viral; Viral Genes; Virus; antitermination factor; bacteriophage T3 RNA polymerase; cell growth regulation; in vivo; inhibitor/antagonist; novel; programs; stem; termination factor; viral DNA

DESCRIPTION (provided by applicant): Our long-term goal is to understand the function and regulation of cellular RNA polymerase (RNAP) in molecular detail. Bacteriophages evolved elaborate mechanisms to regulate transcription of bacterial host to serve viral needs. The number of phage-encoded transcription regulators exceeds the number of bacterial regulators by orders of magnitude. Phage regulatory systems are usually compact, robust and efficient. Studies of a handful of phage-induced modifications of host RNAP provided important paradigms of regulation of gene expression that are applicable to bacteria and higher organisms. The goal of this research is to study phage-induced modifications of bacterial host RNAP and the role of these modifications in viral development. In vitro, RNAP sites that are targeted by phage regulators will be identified and the mechanisms of action of phage regulators will be determined. In vivo, genetic and genomic approaches will be used to understand the biological consequences of RNAP modifications by phage-encoded inhibitors. The following model systems will be analyzed: The T4 phage. Molecular mechanism of termination factor Ale will be studied. The mechanism of negative regulation of host and early viral genes and positive regulation of middle viral genes by T4 AsiA will be determined. The role of ADP-ribosylation of RNAP alpha in regulation of early and middle viral transcription will be studied. The T7 phage. The role of RNAP beta phosphorylation by T7 gp0.7, the molecular mechanism of E. coli RNAP transcription inhibition by T7 gp2, and the role of gp2 in viral DNA packaging will be investigated. The Sp6 phage. Sp6-encoded inhibitor(s) of S. typhimurium RNAP will be purified and characterized. The Xp10 phage. Molecular mechanism of a novel Xp 10-encoded antitermination factor p7 will be identified. Global transcription profiling will be used to better understand gene expression strategy of Xp10, a highly unusual phage that appears to combine the regulatory paradigms of well-studied T7 and lambda phages. The thematic unity of this application stems from its focus on negative regulation of bacterial RNAP by covalent modifications or RNAP-interacting proteins during viral development. Studies of phage-encoded proteins and modifications of host RNAP will lead to deeper understanding of viral biology. On the other hand, phage-encoded transcription inhibitors will be used as molecular probes to better understand RNAP mechanism and regulation and to uncover RNAP sites that can be targets for drug design.

描述（由申请人提供）：我们的长期目标是了解细胞RNA聚合酶（RNAP）的分子细节功能和调控。噬菌体进化出复杂的机制来调节细菌宿主的转录以满足病毒的需要。噬菌体编码的转录调节因子的数量超过了细菌调节因子的数量。噬菌体调节系统通常紧凑、稳健和高效。对少数噬菌体诱导的宿主RNAP修饰的研究提供了适用于细菌和高等生物的基因表达调控的重要范例。本研究的目的是研究噬菌体诱导的细菌宿主RNAP的修饰以及这些修饰在病毒发育中的作用。在体外，将鉴定噬菌体调节剂靶向的RNAP位点，确定噬菌体调节剂的作用机制。在体内，遗传学和基因组学方法将被用来了解通过噬菌体编码抑制剂修饰RNAP的生物学后果。将分析以下模型系统：