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

点击翻译按钮获取中文摘要

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.