DNA Replication in the Gram Positive Bacterium Bacillus Subtilis
革兰氏阳性菌枯草芽孢杆菌中的 DNA 复制
基本信息
- 批准号:0919961
- 负责人:
- 金额:$ 60.43万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:2009
- 资助国家:美国
- 起止时间:2009-09-01 至 2012-08-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5). To date, E. coli serves as the only cellular system for which a complete replication fork can be reconstituted from purified proteins. In this project, a highly divergent bacterium, Baccilus subtilis, will be explored to learn what principles established from the E. coli model are general and what variations can be employed for the rapid, processive, coordinated replication of a chromosome. B. subtilis is an attractive system in that regard. Through genetic, molecular and cell biology work it is known that two distinct replicases are required, unlike the E. coli system where the DNA polymerase III holoenzyme alone suffices. The two-replicase system is reminiscent of eukaryotic systems. It is also known that two proteins are required to load the helicase, unlike E. coli, but perhaps similar to the suspected Orc6/Cdt1 requirement in eukaryotes. B. subtilis also employs two novel proteins, DnaD and DnaB, that have no homologs in E. coli. Thirteen proteins, predicted from genetics, have been expressed and purified and have been used to reconstitute a robust, rolling circle replicative reaction that recapitulates the general features of B. subtilis chromosomal replication in vivo. A rigorous study will be conducted to elucidate the cooperative interactions of replication proteins that participate in the helicase assembly process. The function of the two replicases, DnaE and PolC, will be perturbed to gain fundamental information regarding their function.Completion of the program will lead to an important mechanistic understanding of DNA replication in a diverse model organism. Fundamental knowledge will also serve as a prototype for comparative studies in less tractable eukaryotic systems. The program will increase training opportunities for undergraduate and graduate researchers, including groups underrepresented in science.
该奖项是根据2009年美国复苏和再投资法案(公法111-5)资助的。迄今为止,E.大肠杆菌作为唯一的细胞系统,其完整的复制叉可以从纯化的蛋白质重建。 本计画将探讨一种高度分化的细菌--枯草芽孢杆菌,以了解其从大肠杆菌建立的原理。大肠杆菌模型是通用的,什么样的变化可以用于快速,进行性,协调复制的染色体。 B。在这方面,枯草芽孢杆菌是一种有吸引力的系统。 通过遗传学、分子生物学和细胞生物学的研究,人们知道,与大肠杆菌不同,需要两种不同的复制酶。大肠杆菌系统,其中DNA聚合酶III全酶单独就足够了。双复制酶系统让人想起真核系统。 还已知需要两种蛋白质来装载解旋酶,这与E.大肠杆菌,但可能类似于怀疑Orc 6/Cdt 1的要求,在真核生物。 B。枯草杆菌还利用了两种新的蛋白质,DnaD和DnaB,它们在大肠杆菌中没有同源物。杆菌 从遗传学预测的13种蛋白质已被表达和纯化,并已被用于重建一个强大的滚环复制反应,该反应概括了B的一般特征。枯草芽孢杆菌染色体在体内复制。 将进行严格的研究,以阐明参与解旋酶组装过程的复制蛋白的合作相互作用。 这两种复制酶,DnaE和PolC的功能,将被扰乱,以获得有关其功能的基本信息。该计划的完成将导致在不同的模式生物中DNA复制的重要机制的理解。 基础知识也将作为一个原型,比较研究在不太听话的真核系统。 该计划将增加本科生和研究生研究人员的培训机会,包括科学领域代表性不足的群体。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Charles McHenry其他文献
Charles McHenry的其他文献
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{{ truncateString('Charles McHenry', 18)}}的其他基金
Polymerase Dynamics at the Replication Fork
复制叉的聚合酶动力学
- 批准号:
1329285 - 财政年份:2013
- 资助金额:
$ 60.43万 - 项目类别:
Continuing Grant
Subunit Arrangement within a Replicative Complex
复制复合体内的亚基排列
- 批准号:
9513248 - 财政年份:1996
- 资助金额:
$ 60.43万 - 项目类别:
Continuing Grant
Request for Acquisition of Instrumentation for the Study of Macromolecular Interactions
请求购买用于研究大分子相互作用的仪器
- 批准号:
9419642 - 财政年份:1995
- 资助金额:
$ 60.43万 - 项目类别:
Standard Grant
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