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的多部分蛋白质在一种称为基因表达的活动中起着至关重要的作用。基因表达是重要的，因为它最终有助于生物体的发育和生存。RNAP可以被认为是为遗传信息提供了一种声音，而这种声音本身是沉默的。了解这种声音是如何被放大和嘘的是生物学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必须首先与代表基因的DNA序列结合，并解开DNA序列的配对链，以接近“模板DNA链”，从而“读取”基因中包含的信息。RNAP的几个移动部分，称为“移动的部分”，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”。来自细菌如大肠杆菌的RNAP的活性通常通过细菌RNAP的“移动的部分”来控制。有趣的是，感染和杀死细菌的病毒通常通过制造非常小的蛋白质来实现，这些蛋白质专门用于干扰细菌RNAP的“移动的部分”的正常功能，这些部分有助于DNA结合和DNA解旋活动。尽管对RNAP进行了四十年的研究，但对RNAP的“移动的部分”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“移动的部分”如何有助于基因表达和（ii）细菌病毒衍生的小蛋白如何抑制细菌RNAP。拟议的研究的结果将有重要的影响，在生物和医学研究的几个方面：了解细菌RNAP的“移动的部分”如何有助于基因表达是很重要的，以阐明一般的基因表达的过程和机制方面，因为实验上容易处理的细菌RNAP是一个很好的模型来研究RNAP在复杂的生物体，如人类的基因表达。了解细菌病毒衍生的小蛋白如何抑制细菌RNAP是重要的，因为它们提供了一个新的切入点来评估靶标，并为开发新的药物产生知识基础，这些药物可以抑制细菌RNAP，从而控制细菌感染性疾病的传播。

项目成果

Activity map of the Escherichia coli RNA polymerase bridge helix.

• DOI: 10.1074/jbc.m110.212902
• 发表时间: 2011-04-22
• 期刊: The Journal of biological chemistry
• 作者: Jovanovic M;Burrows PC;Bose D;Cámara B;Wiesler S;Zhang X;Wigneshweraraj S;Weinzierl RO;Buck M
• 通讯作者: Buck M

Opening and closing of the bacterial RNA polymerase clamp.

• DOI: 10.1126/science.1218716
• 发表时间: 2012-08-03
• 期刊: Science (New York, N.Y.)
• 作者: Chakraborty A;Wang D;Ebright YW;Korlann Y;Kortkhonjia E;Kim T;Chowdhury S;Wigneshweraraj S;Irschik H;Jansen R;Nixon BT;Knight J;Weiss S;Ebright RH
• 通讯作者: Ebright RH

Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2.

• DOI: 10.1016/j.molcel.2012.06.013
• 发表时间: 2012-09-14
• 期刊: MOLECULAR CELL
• 影响因子: 16
• 作者: James, Ellen;Liu, Minhao;Sheppard, Carol;Mekler, Vladimir;Camara, Beatriz;Liu, Bing;Simpson, Pete;Cota, Ernesto;Severinov, Konstantin;Matthews, Steve;Wigneshweraraj, Sivaramesh
• 通讯作者: Wigneshweraraj, Sivaramesh

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