Molecular mechanisms of DNA replication

描述(由申请人提供)： 摘要DNA复制是所有生物体在细胞分裂前精确复制遗传物质的基本过程。这个过程的中心是一种解旋酶，它利用ATP水解酶来分离碱基配对的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复合体。在分子水平上，人们对MCMS用于对DNA进行操作的机制知之甚少。这项拟议的研究将通过提供MCM蛋白质与DNA和ATP化合物相互作用的详细图片来填补知识空白。这些化合物将通过一种协调的方法在分子水平上进行研究，包括X射线结晶学的结构研究和体外方法来研究它们的功能和相互作用。该项目已经获得了大量的初步数据，包括MCM：DNA共晶体结构，MCM：ADP的重要初步衍射晶体，纯化的蛋白质，以及来自SPR研究的功能信息。

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.