Initiation of DNA replication in Bacillus subtilis

枯草芽孢杆菌中 DNA 复制的启动

• 批准号: BB/E006450/1
• 负责人: Panos Soultanas
• 金额: $ 45.08万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 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%2FE006450%2F1
• 关键词: Initiation; DNA; replication; Bacillus; subtilis

Reorganising the architecture of the bacterial genetic material is a complicated and dynamic process with crucial consequences to bacterial physiology. Significant reorganisation of the genetic material is required to set the scene for the start of DNA replication and also to assist the separation of the new genetic material from the old one. The molecular mechanisms behind DNA reorganisation and remodelling are not understood. We have recently discovered that two essential replication proteins known as DnaD and DnaB from the bacterium Bacillus subtilis exhibit significant opposing DNA-remodelling activities. In an effort to establish whether these remodelling activities are of direct relevance to DNA replication and to bacterial physiology we are now concentrating our efforts to reveal the molecular mechanisms that underpin these remodelling activities and also to study the effects of limiting these two proteins in the cell. We are aiming to solve the crystal structures of the two domains of DnaD and also of the full-length protein. Mutagenesis methods in parallel with the structural studies but also assisted by the forth-coming structures, together with supercoiling and genetic complementation experiments will be used to study the mechanisms of DNA remodelling and their significance to bacterial physiology. The findings of this study will be crucial if we are to undertand how the genetic material is reorganised in relation to DNA replication in a large class of bacteria known as gram positives. This class of bacteria contains important pathogens like Staphylococcus aureus (responsible for MRSA infections) and our findings will be of general applicability to the discovery of new antibiotic targets.

重组细菌遗传物质的结构是一个复杂而动态的过程，对细菌生理学具有重要影响。需要对遗传物质进行重大重组，以设置DNA复制开始的场景，并帮助新遗传物质与旧遗传物质分离。DNA重组和重塑背后的分子机制尚不清楚。我们最近发现，来自枯草芽孢杆菌的两种称为DnaD和DnaB的必需复制蛋白表现出显著相反的DNA重塑活性。为了确定这些重塑活动是否与DNA复制和细菌生理学直接相关，我们现在正集中精力揭示支持这些重塑活动的分子机制，并研究限制细胞中这两种蛋白质的影响。我们的目标是解决DnaD的两个结构域以及全长蛋白质的晶体结构。与结构研究平行的诱变方法，但也得到了即将到来的结构的帮助，以及超螺旋和遗传互补实验将用于研究DNA重塑的机制及其对细菌生理学的意义。这项研究的发现将是至关重要的，如果我们要了解遗传物质是如何重组与DNA复制在一个大类的细菌称为革兰氏阳性。这类细菌含有重要的病原体，如金黄色葡萄球菌（负责MRSA感染），我们的研究结果将普遍适用于发现新的抗生素靶点。

项目成果

DnaB proteolysis in vivo regulates oligomerization and its localization at oriC in Bacillus subtilis.

• DOI: 10.1093/nar/gkp1236
• 发表时间: 2010-05
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Grainger WH;Machón C;Scott DJ;Soultanas P
• 通讯作者: Soultanas P

RepD-mediated recruitment of PcrA helicase at the Staphylococcus aureus pC221 plasmid replication origin, oriD.

• DOI: 10.1093/nar/gkp1153
• 发表时间: 2010-04
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Machón C;Lynch GP;Thomson NH;Scott DJ;Thomas CD;Soultanas P
• 通讯作者: Soultanas P

Insights into the structure and assembly of the Bacillus subtilis clamp-loader complex and its interaction with the replicative helicase.

• DOI: 10.1093/nar/gkt173
• 发表时间: 2013-05
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Afonso JP;Chintakayala K;Suwannachart C;Sedelnikova S;Giles K;Hoyes JB;Soultanas P;Rafferty JB;Oldham NJ
• 通讯作者: Oldham NJ

Conserved residues of the C-terminal p16 domain of primase are involved in modulating the activity of the bacterial primosome.

• DOI: 10.1111/j.1365-2958.2008.06155.x
• 发表时间: 2008-04
• 期刊: Molecular microbiology
• 影响因子: 3.6
• 作者: Chintakayala K;Larson MA;Griep MA;Hinrichs SH;Soultanas P
• 通讯作者: Soultanas P

Domain swapping reveals that the C- and N-terminal domains of DnaG and DnaB, respectively, are functional homologues.

• DOI: 10.1111/j.1365-2958.2007.05617.x
• 发表时间: 2007-03
• 期刊: Molecular microbiology
• 影响因子: 3.6
• 作者: Chintakayala K;Larson MA;Grainger WH;Scott DJ;Griep MA;Hinrichs SH;Soultanas P
• 通讯作者: Soultanas P