Molecular mechanisms of DNA replication

DNA复制的分子机制

• 批准号: 9336317
• 负责人: Eric Enemark
• 金额: $ 38.2万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336317
• 关键词: ATP phosphohydrolase; ATPase Domain; Affect; Base Pairing; Binding; Biochemical; C-terminal; Cell division; Clinical Trials; Complex; Crystallization; DNA; DNA Binding; DNA Maintenance; DNA Structure; DNA biosynthesis; DNA replication fork; Data; Disease; Elements; Enzymes; Eukaryota; Eukaryotic Cell; Event; Funding; Genetic; Genetic Materials; Goals; Growth; Human; Human Biology; Hydrolysis; In Vitro; Investigation; Knowledge; Lead; Life; MCM6 gene; Maintenance; Malignant Neoplasms; Measures; Mediating; Methods; Molecular; Monitor; Movement; N-terminal; Nucleotides; Organism; Phosphotransferases; Polymerase; Positioning Attribute; Preparation; Process; Proteins; Replication-Associated Process; Research; Rotation; Side; Single-Stranded DNA; Structure; Time; X-Ray Crystallography; cancer therapy; experimental study; helicase; inhibitor/antagonist; insight; interest; pathogen

ABSTRACT DNA replication is a fundamental process for all organisms to precisely duplicate genetic material prior to cell division. Central to the process is a helicase enzyme that uses ATP-hydrolysis to separate base-paired DNA to allow polymerases to gain access to synthesize complementary strands and also to drive the replication machinery along the DNA. In human and other eukaryotic cells, the helicase engine is the 6- protein MCM complex. The mechanism for MCM-mediated manipulation of DNA is poorly understood at the molecular level because of a long-standing lack of structural information for several discrete functional states. The proposed research will fill a knowledge gap by providing detailed pictures of MCM proteins interacting with ATP compounds and with different forms of DNA. These will be studied at the molecular level by a coordinated approach involving structural studies by X-ray crystallography and in vitro methods to study their functions and interactions. A large body of preliminary data has been obtained for this project, including MCM:ATP co-crystal structures, important preliminary diffracting crystals of human MCMs with single-stranded DNA, and several purified proteins with confirmed biochemical activity.

抽象的 DNA复制是所有生物体在形成之前精确复制遗传物质的基本过程。 细胞分裂。该过程的核心是解旋酶，它利用 ATP 水解来分离碱基对 DNA允许聚合酶合成互补链并驱动 沿着DNA的复制机器。在人类和其他真核细胞中，解旋酶引擎是 6- 蛋白质 MCM 复合物。人们对 MCM 介导的 DNA 操纵机制知之甚少。 分子水平，因为长期缺乏几个离散功能的结构信息 州。拟议的研究将通过提供 MCM 蛋白的详细图片来填补知识空白 与 ATP 化合物和不同形式的 DNA 相互作用。这些将在分子水平上进行研究 通过协调方法，包括通过 X 射线晶体学和体外方法进行结构研究， 研究它们的功能和相互作用。该项目已获得大量初步数据， 包括 MCM:ATP 共晶结构、人类 MCM 的重要初步衍射晶体 单链 DNA 和几种具有已证实生化活性的纯化蛋白质。