Mechanism of action of small-molecule inhibitors of bacterial gene transcription.

细菌基因转录小分子抑制剂的作用机制。

• 批准号: BB/E023703/1
• 负责人: Sivaramesh Wigneshweraraj
• 金额: $ 109.05万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE023703%2F1
• 关键词: Mechanism; action; small; molecule; inhibitors

In all living organisms, a multi-part protein, called RNAP, plays a crucial role in an activity of called gene expression. Gene expression is important because it ultimately contributes to the development and survival of the living organism. The RNAP can be regarded as providing a voice to genetic information that, on its own, is silent. Learning how that voice is amplified - and shushed - is a critical stepping stone to many areas of biological and medical research. Thus, the RNAP is a very important protein that directly contributes to the development and survival of all living organisms. To express a gene, the RNAP, must first bind to the DNA-sequence that represents the gene and unwind the paired strands of the DNA-sequence to access the 'template-DNA-strand' to 'read' the information harboured in the gene. Several moving parts of the RNAP, called 'mobile parts' contribute to the DNA binding and DNA unwinding activities of the RNAP, and therefore are often 'control points' for regulating the activity of the RNAP. The activity of the RNAP from bacteria, such as Escherichia coli, is often controlled through the 'mobile parts' of the bacterial RNAP. Intriguingly, viruses which infect and kill bacteria, often do so by making very small proteins, specifically aimed to interfere with the normal functioning of the 'mobile parts' of the bacterial RNAP, which contribute to the DNA binding and DNA unwinding activities. Despite four decades of research on RNAP, very little is known about how the 'mobile parts' of the RNAP contribute to gene expression. The timely work proposed in this application is aimed at studying (i) how two 'mobile parts' of the RNAP contribute to gene expression and (ii) how bacterial virus derived small proteins inhibit the bacterial RNAP. The outcome of the proposed research will have important implications on several aspects of biological and medical research: Understanding how the 'mobile parts' of the bacterial RNAP contribute to gene expression is important to elucidate the process and mechanistic aspects of gene expression in general, since the experimentally easily tractable bacterial RNAP is an excellent model to study gene expression by RNAP in complex organisms, like humans. Understanding how bacterial virus derived small proteins inhibit the bacterial RNAP is significant because they provide a novel point of entry to evaluate targets and generate a knowledge-base for the development of new drugs, which can inhibit the bacterial RNAP and so control the spread of bacterial infectious diseases.

在所有生物体中，一种称为 RNAP 的多部分蛋白质在基因表达活动中发挥着至关重要的作用。基因表达很重要，因为它最终有助于生物体的发育和生存。 RNAP 可以被视为为遗传信息提供了声音，而遗传信息本身是沉默的。了解声音如何被放大和静音是生物和医学研究许多领域的关键垫脚石。因此，RNAP是一种非常重要的蛋白质，直接有助于所有生物体的发育和生存。为了表达基因，RNAP 必须首先与代表该基因的 DNA 序列结合，并解开 DNA 序列的成对链，以访问“模板 DNA 链”，以“读取”基因中蕴含的信息。 RNAP 的几个移动部分，称为“移动部分”，有助于 RNAP 的 DNA 结合和 DNA 解旋活性，因此通常是调节 RNAP 活性的“控制点”。细菌（例如大肠杆菌）的 RNAP 活性通常通过细菌 RNAP 的“移动部分”进行控制。有趣的是，病毒感染并杀死细菌时，通常会制造非常小的蛋白质，专门干扰细菌 RNAP 的“移动部分”的正常功能，而 RNAP 有助于 DNA 结合和 DNA 解旋活动。尽管对 RNAP 的研究已有四十年之久，但人们对 RNAP 的“移动部分”如何促进基因表达知之甚少。本申请中提出的及时工作旨在研究（i）RNAP 的两个“移动部分”如何促进基因表达以及（ii）细菌病毒衍生的小蛋白如何抑制细菌 RNAP。拟议研究的结果将对生物学和医学研究的几个方面产生重要影响：了解细菌 RNAP 的“移动部分”如何促进基因表达对于阐明一般基因表达的过程和机制非常重要，因为实验上易于处理的细菌 RNAP 是研究人类等复杂生物体中 RNAP 基因表达的绝佳模型。了解细菌病毒衍生的小蛋白如何抑制细菌 RNAP 具有重要意义，因为它们提供了一个新的切入点来评估靶标并为开发新药提供知识库，从而抑制细菌 RNAP 从而控制细菌的传播传染病。

项目成果

Overexpression of Escherichia coli udk mimics the absence of T7 Gp2 function and thereby abrogates successful infection by T7 phage.

大肠杆菌 udk 的过度表达模拟 T7 Gp2 功能的缺失，从而消除 T7 噬菌体的成功感染。

• DOI: http://dx.10.1099/mic.0.064527-0
• 发表时间: 2013
• 期刊: Microbiology (Reading, England)
• 作者: Shadrin A

Opening and closing of the bacterial RNA polymerase clamp.

细菌 RNA 聚合酶夹的打开和关闭。

• DOI: http://dx.10.1126/science.1218716
• 发表时间: 2012
• 期刊: Science (New York, N.Y.)
• 作者: Chakraborty A

Sensing DNA opening in transcription using quenchable Förster resonance energy transfer.

使用可淬灭的 Förster 共振能量转移来检测转录中的 DNA 打开。

• DOI: 10.1021/bi101184g
• 发表时间: 2010-10-06
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: T. Cordes;Y. Santoso;Ale;ra I. Tomescu;ra;Kristofer Gryte;L. Hwang;B. Cámara;S. Wigneshweraraj;A. Kapanidis
• 通讯作者: A. Kapanidis

Coupling sigma factor conformation to RNA polymerase reorganisation for DNA melting.

将 Sigma 因子构象与 RNA 聚合酶重组耦合以进行 DNA 熔解。

• DOI: http://dx.10.1016/j.jmb.2009.01.052
• 发表时间: 2009-03-27
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Burrows, Patricia C.;Joly, Nicolas;Cannon, Wendy V.;Camara, Beatriz P.;Rappas, Mathieu;Zhang, Xiaodong;Dawes, Kathleen;Nixon, B. Tracy;Wigneshweraraj, Siva R.;Buck, Martin
• 通讯作者: Buck, Martin

Activity map of the Escherichia coli RNA polymerase bridge helix.

大肠杆菌 RNA 聚合酶桥螺旋的活性图。

• DOI: http://dx.10.1074/jbc.m110.212902
• 发表时间: 2011
• 期刊: The Journal of biological chemistry
• 作者: Jovanovic M