High-resolution analysis of the spatio-temporal control of the DNA replication activation

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The mammalian cell has the challenge to duplicate its entire genome once and only once every time it divides. Our objective is to understand this central cell biological process at the level of the single replication unit and at the level of the whole genome. The basic mechanisms of DNA replication have been established by a powerful combination of genetics and in vitro biochemistry. From this body of work, it is clear that the fundamental features and components of DNA replication are well conserved throughout evolution from bacteria up to mammals. A comprehensive list of proteins that are involved in the process and their activities has been compiled, and it is now possible to provide a relatively detailed description of the DNA replication machinery on a molecular level. However, genomic DNA in eukaryotic cells is hierarchically packed within the nucleus and genome duplication requires the concerted effort of many thousands of individual replication units over time. As such, it is important to understand how DNA replication is regulated in space and time across the entire genome and within the cellular context.The goal of our project is to elucidate the spatio-temporal control of the activation mechanism of DNA replication in mammalian cells. In particular, we will address whether i) DNA higher order structure, ii) DNA modifications and iii) DNA (com)position in the nucleus regulate DNA replication dynamics. The outcome of our project should contribute to, on the one hand, the elucidation of the structure-functional dynamics of the DNA replication process in the cellular context and, on the other hand, provide evidence for its activation mechanism.