Mechanism for loading the E. coli beta clamp on DNA

DNA 上大肠杆菌 beta 钳的加载机制

• 批准号: 7620283
• 负责人: SARAH Katharine SCOUTEN
• 金额: $ 0.87万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-02 至 2008-09-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7620283
• 关键词: Affinity; Binding; Biochemical; Biological Assay; Biological Models; C-terminal; Cells; Cleaved cell; Complex; Conflict (Psychology); DNA; DNA Damage; DNA Repair; DNA biosynthesis; Data; Escherichia coli; Eukaryota; Eukaryotic Cell; Excision; Fellowship; Genome; Goals; Human; Individual; Learning; Life; Literature; Malignant Neoplasms; Mediating; Modeling; Numbers; Organism; Play; Positioning Attribute; Prokaryotic Cells; Proteins; Relative (related person); Reporting; Research; Role; Site; Structure; Surface; Tail; Virus; Work; base; chromosome replication; dimer; human disease; insight; monomer; mutant; prevent; repaired; stoichiometry

The long term goal of this research is to understand how organisms coordinate the replication and repair of their genomes. The use of processivity clamps that must be loaded onto DNA during replication of chromosomes by an ATP dependent clamp loader complex is highly conserved by all forms of life. Understanding how organisms coordinate DNA replication and repair is the fundamental basis for understanding human diseases such as cancer. The goal of the work proposed here is to better define the mechanism of clamp loading in the E. coli model system. The E. coli clamp loader (y complex) is a multisubunit complex that undergoes a conformational change upon binding ATP which facilitates opening of the p clamp and its subsequent loading onto DNA. The contacts of the individual clamp loader subunits with P differ between the ATP bound and free states of y complex. The interactions of the y complex subunits with p in both the presence and absence of ATP will be determined. Biochemical assays using various clamp and clamp loader mutants will be uilized to accomplish this. The insight gained from E. coli can provide a framework for elucidating the mechanisms of eukaryotes, including humans.

这项研究的长期目标是了解生物如何协调复制和修复 他们的基因组。复制过程中必须将其加载到DNA上的加工性夹具的使用 ATP依赖性夹具装载机复合物的染色体由所有形式的生命都高度保守。 了解生物如何协调DNA复制和维修是基本的基础 了解人类疾病，例如癌症。这里提出的工作的目的是更好地定义 大肠杆菌模型系统中的夹具载荷的机理。大肠杆菌夹具装载机（Y配合物）是 多育种复合物在结合ATP时经历构象变化，这有助于开放 P夹具及其随后的加载到DNA上。单个夹具装载机亚基与 p在Y复合物的ATP结合和自由状态之间有所不同。 Y复合体亚基的相互作用 在ATP的存在和不存在的情况下，将确定P。使用各种生化测定 夹具和夹具装载机突变体将被尿e以实现这一目标。从大肠杆菌获得的洞察力可以 提供一个阐明包括人类在内的真核生物机制的框架。