High-Resolution Mutagenesis of the 'Bridge Helix' and 'Switch 1' Domains of Archaeal RNA Polymerase

古细菌 RNA 聚合酶“桥螺旋”和“开关 1”结构域的高分辨率诱变

• 批准号: BB/D00862X/1
• 负责人: Robert Weinzierl
• 金额: $ 30.55万
• 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: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD00862X%2F1
• 关键词: Resolution; Mutagenesis; Bridge; Helix; Switch

The majority of cell types contain the same genetic material to encode the various cellular components. Differences in cell types (e.g. neurons as compared to muscle cells) arise because different parts of the genetic material are read out. RNA polymerases (RNAPs) are a crucial part of the cellular machinery involved in the selective reading mechanism (transcription) and it is therefore important to understand their molecular structure and function in considerable detail. By studying the chemical and physical mechanisms underlying RNAP function we can begin to understand the way these mechanisms are controlled within the cell to reflect the different requirements for gene expression that are posed by various metabolic, developmental and disease states. In this project we intend to study two key structures that are involved directly in the function of RNAP to obtain an insight into the underlying molecular mechanisms.

大多数细胞类型含有相同的遗传物质来编码各种细胞成分。细胞类型的差异（例如神经元与肌肉细胞相比）是因为遗传物质的不同部分被读出。RNA聚合酶（RNAPs）是参与选择性阅读机制（转录）的细胞机器的关键部分，因此相当详细地了解它们的分子结构和功能是重要的。通过研究RNAP功能的化学和物理机制，我们可以开始了解这些机制在细胞内的控制方式，以反映各种代谢、发育和疾病状态对基因表达的不同要求。在这个项目中，我们打算研究直接参与RNAP功能的两个关键结构，以深入了解潜在的分子机制。

项目成果

Bridge helix and trigger loop perturbations generate superactive RNA polymerases.

• DOI: 10.1186/jbiol98
• 发表时间: 2008-12-02
• 期刊: Journal of biology
• 作者: Tan L;Wiesler S;Trzaska D;Carney HC;Weinzierl RO
• 通讯作者: Weinzierl RO

Construction and functional analyses of a comprehensive sigma54 site-directed mutant library using alanine-cysteine mutagenesis.

使用丙氨酸-半胱氨酸诱变构建综合 sigma54 定点突变体库并进行功能分析。

• DOI: 10.1093/nar/gkp419
• 发表时间: 2009
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Xiao Y

Nanomechanical constraints acting on the catalytic site of cellular RNA polymerases.

作用于细胞 RNA 聚合酶催化位点的纳米力学约束。

• DOI: 10.1042/bst0380428
• 发表时间: 2010
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Weinzierl RO

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

The Bridge Helix of RNA polymerase acts as a central nanomechanical switchboard for coordinating catalysis and substrate movement.

• DOI: 10.1155/2011/608385
• 发表时间: 2011
• 期刊: Archaea (Vancouver, B.C.)
• 作者: Weinzierl RO