Investigating the DnaA-trio, a new essential bacterial replication origin element that specifies single-stranded DNA initiator binding

研究 DnaA-trio，一种新的必需细菌复制起点元件，可指定单链 DNA 起始子结合

• 批准号: BB/P018432/1
• 负责人: Heath Murray
• 金额: $ 44.48万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP018432%2F1
• 关键词: Investigating; DnaA; trio; new; essential

In all cells genome duplication requires key proteins ("initiators") to unwind the DNA double helix at specific sites ("origins"). DNA duplex unwinding provides the replication machinery access to single strands, which act as templates for new rounds of DNA synthesis. While initiators are highly conserved throughout all organisms and well characterised, origins are enigmatic. In fact within higher organisms, such as humans, we have yet to define what constitutes an origin. Bacteria, with their relatively simple and well characterised structure and physiology, are ideal systems with which to study the molecular mechanisms of DNA replication because they are readily amenable to genetic manipulation and their proteins tend to be tractable subjects for biochemical and structural analyses. Recently we reported the identification of a novel replication origin element (termed the "DnaA-trio") and showed that it acts to promote single-strand DNA binding activity of the master bacterial replication initiator protein, DnaA. We proposed that this essential recognition element represents a conserved component of the core bacterial chromosome origin. One of the goals of this proposal is to determine how widespread the DnaA-trio is within the bacterial kingdom. We are particularly interested in testing whether this chromosome origin element is present and active in human pathogens such as Staphylococcus aureus and Helicobacter pylori, since this would identify a novel target for antibiotic development. Another goal is to characterize the interaction of DnaA-trios with the initiator protein DnaA. Further understanding of the molecular mechanisms underlying DnaA recognition and activity with this new chromosome origin element will provide cutting-edge knowledge regarding a fundamental biological process that is essential for viability and proliferation. It is important to stress that all initiator proteins throughout the three kingdoms of life contain a related protein fold (the initiator specific AAA+ motif), indicating that they evolved from a common ancestor and that they share common activities. Therefore, the findings from this research project will inform how eukaryotic initiator proteins act and will underpin the search for replication origin elements in higher organisms, which at this moment are ill defined.

在所有细胞中，基因组复制都需要关键蛋白（“引发剂”）来在特定位点（“起源”）解开 DNA 双螺旋。 DNA 双链体解旋为复制机制提供了接触单链的途径，单链充当新一轮 DNA 合成的模板。虽然引发剂在所有生物体中高度保守且特征明确，但其起源却是个谜。事实上，在人类等高等生物体内，我们尚未定义起源的构成。细菌具有相对简单且特征明确的结构和生理学，是研究 DNA 复制分子机制的理想系统，因为它们很容易进行基因操作，并且它们的蛋白质往往是生化和结构分析的易处理对象。最近，我们报道了一种新型复制起点元件（称为“DnaA-trio”）的鉴定，并表明它可以促进主细菌复制起始蛋白 DnaA 的单链 DNA 结合活性。我们提出，这种重要的识别元件代表了核心细菌染色体起源的保守成分。该提案的目标之一是确定 DnaA-三重奏在细菌王国中的分布范围。我们特别感兴趣的是测试这种染色体起源元件是否存在于金黄色葡萄球菌和幽门螺杆菌等人类病原体中并具有活性，因为这将为抗生素开发确定一个新靶点。另一个目标是表征 DnaA-trios 与起始蛋白 DnaA 的相互作用。进一步了解这种新染色体起源元件的 DnaA 识别和活性的分子机制，将提供关于生存和增殖所必需的基本生物过程的前沿知识。需要强调的是，生命的三个王国中的所有起始蛋白都包含相关的蛋白折叠（起始特定的 AAA+ 基序），这表明它们是从共同的祖先进化而来的，并且具有共同的活性。因此，该研究项目的发现将告知真核起始蛋白如何发挥作用，并将支持在高等生物体中寻找复制起始元件，目前这些元件尚不清楚。

项目成果

The DNA replication initiation protein DnaD recognises a specific strand of the Bacillus subtilis chromosome origin

DNA 复制起始蛋白 DnaD 识别枯草芽孢杆菌染色体起源的特定链

• DOI: http://dx.10.25418/crick.23132663
• 发表时间: 2023
• 作者: Winterhalter C

Identification of a basal system for unwinding a bacterial chromosome origin.

鉴定用于解开细菌染色体起源的基础系统。

• DOI: http://dx.10.15252/embj.2019101649
• 发表时间: 2019
• 期刊: The EMBO journal
• 作者: Richardson TT

SirA inhibits the essential DnaA:DnaD interaction to block helicase recruitment during Bacillus subtilis sporulation.

SirA 抑制必需的 DnaA:DnaD 相互作用，以阻止枯草芽孢杆菌孢子形成过程中解旋酶的募集。

• DOI: http://dx.10.1093/nar/gkac1060
• 发表时间: 2023
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Winterhalter C

The DNA replication initiation protein DnaD recognises a specific strand of the Bacillus subtilis chromosome origin.

DNA 复制起始蛋白 DnaD 识别枯草芽孢杆菌染色体起点的特定链。

• DOI: http://dx.10.1093/nar/gkad277
• 发表时间: 2023
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Winterhalter C

SirA inhibits the essential DnaA:DnaD interaction to block helicase recruitment during Bacillus subtilis sporulation

SirA 抑制必需的 DnaA:DnaD 相互作用，以阻止枯草芽孢杆菌孢子形成过程中解旋酶的招募

• DOI: http://dx.10.1101/2022.04.18.488658
• 发表时间: 2022
• 作者: Winterhalter C