Molecular mechanisms of DNA replication

DNA复制的分子机制

基本信息

批准号：
8858639
负责人：
Eric Enemark
金额：
$ 33.25万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8858639
关键词：
ATP Hydrolysis ATP phosphohydrolase ATPase Domain Address Baculoviruses Base Pairing Binding Biochemical Biological Biological Models Cell division Cells Clinical Trials Complex DNA DNA Binding DNA Maintenance DNA biosynthesis Data Defect Disease Drug Design Enzymes Eukaryota Eukaryotic Cell Event Face Genetic Genetic Materials Goals Growth Human Human Biology Image In Vitro Insecta Investigation Knowledge Lead Length Life MCM2 gene Maintenance Malignant Neoplasms Mediating Methods Modeling Molecular Movement Mutagenesis N-terminal Nucleotides Oligonucleotides Oligosaccharides Organism Phosphotransferases Polymerase Preparation Process Protein Binding Proteins Reagent Recombinants Reporting Research Single-Stranded DNA Structure Structure-Activity Relationship Techniques Work X ray diffraction analysis X-Ray Crystallography X-Ray Diffraction base cancer therapy cofactor ds-DNA helicase human MCM2 protein inhibitor/antagonist insight interest pathogen reconstitution tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): 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 utilizes ATP-hydrolysis to separate base- paired DNA to allow polymerases to gain access to synthesize complementary strands as well as to drive the replication machinery along the DNA. In human and other eukaryotic cells, the helicase engine is the hexameric MCM complex. The mechanisms that MCMs use to operate upon DNA are poorly understood at the molecular level. The proposed research will fill a knowledge gap by providing detailed pictures of MCM proteins interacting with DNA and ATP compounds. 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 considerable body of preliminary data has been obtained for this project that includes MCM:DNA co-crystal structures, important preliminary diffracting crystals of MCM:ADP, purified proteins, and functional information from SPR studies.

描述（由申请人提供）：摘要 DNA 复制是所有生物体在细胞分裂之前精确复制遗传物质的基本过程。该过程的核心是解旋酶，它利用 ATP 水解来分离碱基配对的 DNA，使聚合酶能够合成互补链并沿着 DNA 驱动复制机制。在人类和其他真核细胞中，解旋酶引擎是六聚体 MCM 复合物。 MCM 用于作用于 DNA 的机制在分子水平上知之甚少。拟议的研究将通过提供 MCM 蛋白与 DNA 和 ATP 化合物相互作用的详细图片来填补知识空白。这些将通过协调方法在分子水平上进行研究，包括通过 X 射线晶体学和体外方法进行结构研究，以研究其功能和相互作用。该项目已获得大量初步数据，包括 MCM:DNA 共晶结构、MCM:ADP 的重要初步衍射晶体、纯化的蛋白质以及 SPR 研究的功能信息。