Abstract
This review discusses recent advances in single-particle cryo-EM and single-molecule approaches used to visualise eukaryotic DNA replication reactions reconstituted in vitro. We comment on the new challenges facing structural biologists, as they turn to describing the dynamic cascade of events that lead to replication origin activation and fork progression.
Highlights
Accurate transfer of genetic information from parental to daughter cells requires that chromosome replication is finely tuned, so that DNA is faithfully copied only once per cell cycle [1]
The MCM helicase is loaded onto replication start sites, during a process known as licensing that occurs in the G1 phase of the cell cycle
Helicase loading requires ATP hydrolysis by MCM [7,8], which prompts the formation of a double hexameric MCM ring encircling duplex DNA [4,9,10,11]
Summary
Accurate transfer of genetic information from parental to daughter cells requires that chromosome replication is finely tuned, so that DNA is faithfully copied only once per cell cycle [1]. At this stage single-stranded DNA becomes exposed and serves as a template for the replicative polymerases, which dynamically associate with the replisome during fork progression [3,16]. We review recent biochemical, single-molecule and structural studies on in vitro reconstituted reactions that recapitulate DNA replication at cellular rates.
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