We show that the human double hexamer engages DNA differently from the yeast double hexamer and generates approximately five base pairs of underwound DNA at the interface between hexamers.
Our work presents a detailed view of how double hexamers are assembled in an organism that uses sequence-independent replication origins, providing evidence for diversity in eukaryotic double hexamer assembly mechanisms.
Unlike in yeast, the ORC6 subunit of the ORC is not essential for initial MCM recruitment or hDH loading, indicating a distinct assembly pathway in humans.
This research represents a first step towards reconstitution of DNA replication initiation with purified human proteins, shedding light on the mechanistic differences in eukaryotic systems.
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