Molecular mechanisms of archaeal DNA replication: replication initiation and the replicative helicase MCM

这项研究计划的重点是嗜热模式古菌硫化叶菌的DNA复制研究。我们将遵循两个主要的方法：第一，研究的复制起始的Orc 1/Cdc 6蛋白，其次表征的假定复制解旋酶MCM。我们将以生物化学的方式研究所涉及的复制酶，并通过详细分析它们的DNA结合特性，它们的酶特性以及最重要的是它们如何以及是否参与较大的蛋白质复合物以分别在起点和复制叉实现其高度调节和协调的细胞功能来分析它们的酶活性。拟议的研究将使我们深入了解在分子水平上的启动和复制机制，并将开辟途径的结构以及单分子研究的热稳定性和动态酶复合物。由于古细菌DNA复制与真核生物DNA复制相似，但复杂性较低，因此计划中的研究也将有助于加深我们对真核生物DNA复制分子机制的理解。

The focus of this research proposal is the study of DNA replication of the thermophilic model archaeon Sulfolobus. We will follow two major approaches: first, study of the replication initiation by the Orc1/Cdc6 proteins and secondly characterization of the putative replicative helicase MCM. We will study the involved replication enzymes biochemically and analyze their enzymatic activities by a detailed analysis of their DNA binding properties, their enzymatic characteristics and most importantly how and whether they are engaged in larger protein complexes to fulfill their highly regulated and coordinated cellular functions at the origin and the replication fork, respectively. The proposed research will give us an insight into the machinery of initiation and replication on a molecular level and shall open up the avenue for structural as well as single-molecule studies of the thermostable and dynamic enzyme complexes involved. Since the archaeal DNA replication is similar to the eukaryal replication although less complex, the planned research will also help to deepen our understanding on the molecular mechanism of eukaryal DNA replication.