Structure and function of replication-hyperstructures in Escherichia coli

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DNA replication is the process by which all living organisms multiply their genetic information as a prerequisite to reproduce. The bacterium Escherichia coli serves as an important model system for investigations of the basic principles underlying DNA replication. The circular E. coli chromosome is replicated bidirectionally from a unique locus, oriC. Replication cycles can overlap resulting in several copies of oriC per cell. Coordination of the ongoing replication at different chromosome sites and the synchronous initiation of all oriCs presents a difficult task for the cell. The proposed research project aims to understand the role of a physical colocalisation of components involved in DNA replication and its regulation. The starting point is the observation that the DNA-binding protein SeqA can form higher order complexes at the oriC and at the replication fork. The actual composition and function of the SeqA containing structure, also termed a hyperstructure, is unknown and subject of this application. Isolation and characterization of proteins and DNA interacting with SeqA and localization of the components in the cell, as well as characterization of mutant strains, will help to understand the function of hyperstructures in chromosome replication.