MECHANISM OF TERMINATION OF DNA REPLICATION

DNA复制终止机制

• 批准号: 2186841
• 负责人: DEEPAK BASTIA
• 金额: $ 26.94万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2186841
• 关键词: Bacillus subtilis; DNA binding protein; DNA replication; Escherichia coli; X ray crystallography; bacterial proteins; cell cycle; chimeric proteins; circular dichroism; computer simulation; crosslink; crystallization; enzyme linked immunosorbent assay; fluorescence microscopy; fluorescence spectrometry; fungal genetics; gene expression; helicase; intermolecular interaction; molecular cloning; mutant; nucleoproteins; protein purification; protein structure function; site directed mutagenesis

The study of the mechanism of regulation of DNA replication and its coordination wit cell division is of obvious importance proliferation such as cancer. Am important, but incompletely understood step of this larger problem is the mechanism and control of replication terminating, and its possible link to cell division. Utilizing E. coli and B. subtilis and the drug resistance plasmid R6K as model systems, we propose to study the mechanism of action and regulation of the terminator proteins. Specifically, we propose to investigate the molecular basis of the polarity of the contrahelicase activities of ter proteins by combined genetic and biochemical approaches. We have recently discovered that the activity of the ter protein is modulated by specific protein-protein interaction with a novel antiterminator protein. We propose to clone, overproduce and purify to homogeneity the anti-ter protein, and to study the biochemistry and physiological consequences of its interaction with the ter protein. Finally, we have crystallized the ter beta protein (RTP) of B. subtilis, and the crystals diffract to a resolution of 2.5A. We propose to solve not only the structure of the apoprotein but also that of the protein-DNA complex.

DNA复制调控机制及其研究 细胞分裂的协调对于增殖具有明显的重要性 例如癌症。 这一步很重要，但不完全理解 更大的问题是复制终止的机制和控制， 及其与细胞分裂的可能联系。 利用大肠杆菌和 B. 枯草芽孢杆菌和耐药质粒 R6K 作为模型系统，我们建议 研究终止子的作用机制和调控 蛋白质。 具体来说，我们建议研究分子基础 ter 蛋白反解旋酶活性的极性 结合遗传和生化方法。 我们最近发现ter蛋白的活性是 通过特定的蛋白质-蛋白质相互作用与新型 抗终止子蛋白。 我们建议克隆、过量生产和纯化 抗蛋白的同质性，并研究其生物化学和 它与ter蛋白相互作用的生理后果。 最后，我们结晶了枯草芽孢杆菌的ter beta蛋白（RTP）， 晶体衍射分辨率为2.5A。 我们建议解决 不仅包括脱辅基蛋白的结构，还包括蛋白质-DNA 的结构 复杂的。