Bacteriophage-derived Inhibitors of the Bacterial RNA Polymerase

噬菌体衍生的细菌 RNA 聚合酶抑制剂

• 批准号: BB/K000233/1
• 负责人: Sivaramesh Wigneshweraraj
• 金额: $ 79.53万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FK000233%2F1
• 关键词: Bacteriophage; derived; Inhibitors; Bacterial; RNA

Many activities that allow organisms to grow, remain alive and viable rely on molecules resembling small machines. In bacteria, the machine which is responsible for accessing information contained in the genes is the RNA polymerase (RNAp), and the process by which the RNAp accesses the information contained in the genes is collectively called gene expression. Therefore, since the RNAp carries out an essential function necessary for the survival of the bacterial cell, the RNAp is also an attractive target for drugs to treat bacterial infections. For instance, the clinically widely used antibiotic rifampicin inhibits the RNAp. However, the frequent emergence of rifampicin-resistant harmful bacteria necessitates research into new compounds to combat bacterial infections by inhibiting the RNAp. Our research focuses on naturally-occurring small proteins which are potent inhibitors of the RNAp and display several of the desirable features for potential development into novel antibiotic-like compounds for inhibiting the RNAp. Here, we will study precisely how three such naturally-occurring RNAp inhibitors, called Gp2, P7 and Gp5.7 inhibit the RNAp from the bacterium Escherichia coli (for Gp2 and Gp5.7) and Xanthomonas orzyae (a bacterial pathogen that causes rice blight). We will employ structural biology to unravel how Gp2, Gp5.7 and P7 look like and interact with the RNAp. We will employ state-of-the art biochemistry and molecular biology to study how Gp2, Gp5.7 and P7 inhibit the RNAp. We will employ a technique called "next-generation sequencing" to study the impact of Gp2 on RNAp activity, i.e. gene expression, at a whole-cell level. In summary, our results will (1) provide novel insights into how the activity of the bacterial RNAp can be controlled, in this case inhibited, by non-bacterial and naturally-occurring factors; (2) establish a solid framework and industrial links for development of novel antibiotic-like lead compounds targeting the bacterial RNAp and (3) provide useful resources for synthetic biology applications.

许多使生物体生长、存活和存活的活动都依赖于类似小型机器的分子。在细菌中，负责访问基因中包含的信息的机器是RNA聚合酶（RNAp），而RNAp访问基因中包含的信息的过程统称为基因表达。因此，由于RNAp执行细菌细胞存活所必需的基本功能，因此RNAp也是治疗细菌感染的药物的有吸引力的靶标。例如，临床上广泛使用的抗生素利福平抑制RNA β。然而，利福平耐药性有害细菌的频繁出现需要研究新的化合物，以通过抑制RNA β来对抗细菌感染。我们的研究重点是天然存在的小蛋白，它们是RNA β的有效抑制剂，并显示出几种理想的功能，可能开发成抑制RNA β的新型类阿片化合物。在这里，我们将精确地研究三种这样的天然存在的RNAp抑制剂，称为Gp 2，P7和Gp5.7，如何抑制来自细菌大肠杆菌（Gp 2和Gp5.7）和黄单胞菌（导致水稻枯萎病的细菌病原体）的RNAp。我们将采用结构生物学来揭示Gp 2，Gp5.7和P7是如何与RNAp相互作用的。我们将采用最先进的生物化学和分子生物学来研究Gp 2，Gp5.7和P7如何抑制RNA β。我们将采用一种称为“下一代测序”的技术，在全细胞水平上研究Gp 2对RNAp活性（即基因表达）的影响。总之，我们的研究结果将（1）为如何通过非细菌和天然存在的因子控制（在这种情况下抑制）细菌RNAp的活性提供新的见解;（2）为开发靶向细菌RNAp的新型类抗生素先导化合物建立坚实的框架和工业联系;（3）为合成生物学应用提供有用的资源。

项目成果

Nitrogen stress response and stringent response are coupled in Escherichia coli.

• DOI: 10.1038/ncomms5115
• 发表时间: 2014-06-20
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Brown, Daniel R.;Barton, Geraint;Pan, Zhensheng;Buck, Martin;Wigneshweraraj, Sivaramesh
• 通讯作者: Wigneshweraraj, Sivaramesh

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

The Xp10 Bacteriophage Protein P7 Inhibits Transcription by the Major and Major Variant Forms of the Host RNA Polymerase via a Common Mechanism.

• DOI: 10.1016/j.jmb.2016.08.004
• 发表时间: 2016-10-09
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Brown, D. R.;Sheppard, C. M.;Burchell, L.;Matthews, S.;Wigneshweraraj, S.
• 通讯作者: Wigneshweraraj, S.

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