MECHANISM OF TERMINATION OF DNA REPLICATION

DNA复制终止机制

• 批准号: 2521755
• 负责人: DEEPAK BASTIA
• 金额: $ 32.36万
• 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 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2521755
• 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; transcription termination

DESCRIPTION (Adapted from applicant's abstract): The molecular mechanism of polar arrest of replication forks, replicative helicases and RNA chain elongation by the bacterial replication terminator proteins will be investigated using x-ray crystallography, mutagenesis and biochemical analysis of the mutant forms of the terminator proteins. The experiments should provide definitive evidence as to whether not only DNA protein interaction between the terminus DNA and the terminator protein but also protein-protein contacts between the arresting and the arrested protein contributed to replication termination. The physiological role of a terminator protein as checkpoints of replication will be investigated by molecular cloning and in vivo analysis. A genetic selection scheme will be used to attempt to clone and study genes from eukaryotic cells that encode replication fork-arresting protein. X-ray crystallography will be use to determine the structure of a terminus-terminator protein complex.

描述（改编自申请人的摘要）： 的分子机制 复制叉、复制解旋酶和 RNA 链的极性停滞 细菌复制终止蛋白的延伸将是 使用 X 射线晶体学、诱变和生物化学进行研究 终止子蛋白突变形式的分析。 实验 应该提供明确的证据证明是否不仅仅是 DNA 蛋白质 末端DNA和终止子蛋白之间的相互作用 阻滞蛋白和被阻滞蛋白之间的蛋白质-蛋白质接触 导致复制终止。 A的生理作用 作为复制检查点的终止子蛋白将通过以下方式进行研究 分子克隆和体内分析。 遗传选择方案将是 用于尝试克隆和研究真核细胞编码的基因 复制叉阻滞蛋白。 X射线晶体学将用于 确定末端-终止子蛋白复合物的结构。