Gene Control in Infection and Lysogeny by Phage Lambda

噬菌体 Lambda 对感染和溶源的基因控制

• 批准号: 8225361
• 负责人: JEFFREY W ROBERTS
• 金额: $ 57.05万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8225361
• 关键词: ATP Hydrolysis; Affect; Bacteria; Bacteriophage lambda; Bacteriophages; Behavior; Biochemical Genetics; Biological Assay; Biological Models; Complement; Complex; Coupled; DNA; DNA Repair; DNA Structure; DNA repair protein; DNA-Directed RNA Polymerase; Disease; Elements; Elongation Factor; Genes; Genetic; Genetic Transcription; Genome; HIV; HIV Infections; Health; Infection; Learning; Lysogeny; Mediating; Modification; Molecular; Molecular Conformation; Mutation; Names; Nature; Pathogenesis; Pathway interactions; Phase; Process; Proteins; RNA; RNA chemical synthesis; Recruitment Activity; Regulation; Regulatory Pathway; Role; Site; Structure; System; Transcript; Transcription Elongation; Transcription Process; Transcriptional Regulation; Work; antitermination; base; design; genetic analysis; in vivo; insight; prevent; transcription termination

DESCRIPTION (provided by applicant): The objective of this project is to understand control of the elongation phase of RNA synthesis by RNA polymerase, a critical facet of transcription regulation. We will utilize a specific antiterminator protein, the Q protein of bacteriophage lambda, which becomes a subunit of RNA polymerase at a genome-specific site and thereby allows expression of downstream genes by preventing transcription termination. This protein provides a highly defined and accessible system with which to obtain mechanistic insights into universal enzymatic processes that control transcription elongation; for example, the TAT protein of the HIV virus is a regulator of transcription elongation that acts in many ways like lambda Q protein and other bacterial antiterminators. A detailed understanding of these mechanisms will allow approaches to therapies that depend upon the design of specific molecular agents. We will learn the nature of a structural modification, named a barrier that lambda Q protein induces in RNA polymerase in order to make it insensitive to terminators. We will study the role of the transcription elongation factor NusA in constructing the barrier, through biochemical and genetic analysis. We will use mutational analysis to discover the sites and pathways of modification of RNA polymerase by lambda Q protein, in particular to understand how Q regulates transcription pausing, a universal functional behavior of RNA polymerase. This work will complement our continuing efforts to understand the mechanism of termination itself. We also will study the mechanism of action of the protein Mfd, which mediates the process of transcription-coupled DNA repair, and acts to dissociate stalled elongation complexes as it recruits DNA repair proteins. Understanding how the energy of ATP is used by Mfd will illuminate the mechanism of termination and the energetic barriers involved in termination. PUBLIC HEALTH RELEVANCE Using bacterial and bacteriophage model systems, this project contributes to understanding basic mechanisms of genetic regulation, an undertaking essential to discovering the molecular basis of disease. It also directly investigates regulatory pathways related to those essential to pathogenesis by HIV and infection by toxic bacteria.

描述（由申请人提供）：本项目的目的是了解RNA聚合酶对RNA合成延伸阶段的控制，这是转录调控的一个关键方面。我们将利用一种特异性的抗终止子蛋白，即λ噬菌体的Q蛋白，它在基因组特异性位点成为RNA聚合酶的亚基，从而通过阻止转录终止来允许下游基因的表达。这种蛋白质提供了一个高度定义和可访问的系统，通过该系统可以获得对控制转录延伸的通用酶促过程的机械见解;例如，HIV病毒的达特蛋白是转录延伸的调节因子，其在许多方面与λ Q蛋白和其他细菌抗终止子一样起作用。对这些机制的详细理解将允许依赖于特定分子试剂的设计的治疗方法。我们将学习一种结构修饰的性质，称为λ Q蛋白在RNA聚合酶中诱导的屏障，以使其对终止子不敏感。我们将通过生物化学和遗传分析研究转录延伸因子NusA在构建屏障中的作用。我们将使用突变分析来发现λ Q蛋白修饰RNA聚合酶的位点和途径，特别是了解Q如何调节转录暂停，这是RNA聚合酶的一种普遍功能行为。这项工作将补充我们为了解终止机制本身所作的持续努力。我们还将研究蛋白质Mfd的作用机制，Mfd介导转录偶联DNA修复的过程，并在招募DNA修复蛋白时解离停滞的延伸复合物。了解ATP的能量如何被Mfd使用将阐明终止机制和终止中涉及的能量障碍。 公共卫生相关性利用细菌和噬菌体模型系统，该项目有助于了解遗传调控的基本机制，这是发现疾病分子基础的一项重要任务。它还直接研究与HIV发病机制和有毒细菌感染相关的调节途径。

项目成果

The phage lambda gene Q transcription antiterminator binds DNA in the late gene promoter as it modifies RNA polymerase.

噬菌体 lambda 基因 Q 转录抗终止子在修饰 RNA 聚合酶时与晚期基因启动子中的 DNA 结合。

• DOI: 10.1016/0092-8674(92)90639-t
• 发表时间: 1992
• 期刊: Cell
• 影响因子: 64.5
• 作者: Yarnell,WS;Roberts,JW
• 通讯作者: Roberts,JW

Specificity and mechanism of antitermination by Q proteins of bacteriophages lambda and 82.

噬菌体 lambda 和 82 的 Q 蛋白抗终止的特异性和机制。

• DOI: 10.1016/0022-2836(89)90122-8
• 发表时间: 1989
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Yang,XJ;Goliger,JA;Roberts,JW
• 通讯作者: Roberts,JW

RecA protein--promoted lambda repressor cleavage: complementation between RecA441 and RecA430 proteins in vitro.

RecA 蛋白——促进 lambda 阻遏蛋白裂解：体外 RecA441 和 RecA430 蛋白之间的互补。

• DOI: 10.1007/bf00328696
• 发表时间: 1984
• 期刊: Molecular & general genetics : MGG
• 作者: Moreau,PL;Roberts,JW
• 通讯作者: Roberts,JW

A potential stem-loop structure and the sequence CAAUCAA in the transcript are insufficient to signal rho-dependent transcription termination at lambda tR1.

转录本中潜在的茎环结构和序列 CAAUCAA 不足以发出 lambda tR1 处 rho 依赖性转录终止的信号。

• DOI: 10.1093/nar/12.2.1287
• 发表时间: 1984
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Lau,LF;Roberts,JW;Wu,R;Georges,F;Narang,SA
• 通讯作者: Narang,SA

Two transcription pause elements underlie a σ70-dependent pause cycle.

两个转录暂停元件构成了依赖于 70 的暂停周期。

• DOI: 10.1073/pnas.1512986112
• 发表时间: 2015
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Strobel,EricJ;Roberts,JeffreyW
• 通讯作者: Roberts,JeffreyW