
ABSTRACT High-resolution structures have not been reported for replicative helicases at a replication fork at atomic resolution, a prerequisite to understand the unwinding mechanism. The eukaryotic replicative CMG helicase contains a Mcm2-7 motor ring, with the N-tier ring in front and the C-tier motor ring behind. The N-tier ring is structurally divided into a zinc finger (ZF) sub-ring followed by the OB fold ring. Here we report the cryo-EM structure of CMG on forked DNA at 3.9 Å, revealing that parental DNA enters the ZF sub-ring and strand separation occurs at the bottom of the ZF sub-ring, where the lagging strand is blocked and diverted sideways by OB hairpin-loops of Mcm3, Mcm4, Mcm6, and Mcm7. Thus, instead of employing a separation pin, unwinding is achieved via a “dam-and-diversion tunnel” for steric exclusion unwinding. The C-tier motor ring contains spirally configured PS1 and H2I loops of Mcms 2, 3, 5, 6 that translocate on the spirally-configured leading strand, and thereby pull the preceding DNA segment through the diversion tunnel for strand separation.
DNA Replication, Saccharomyces cerevisiae Proteins, Science, Q, Minichromosome Maintenance Complex Component 3, Cell Cycle Proteins, Saccharomyces cerevisiae, Minichromosome Maintenance Complex Component 7, Minichromosome Maintenance Complex Component 6, Article, Minichromosome Maintenance Complex Component 4, DNA, Fungal
DNA Replication, Saccharomyces cerevisiae Proteins, Science, Q, Minichromosome Maintenance Complex Component 3, Cell Cycle Proteins, Saccharomyces cerevisiae, Minichromosome Maintenance Complex Component 7, Minichromosome Maintenance Complex Component 6, Article, Minichromosome Maintenance Complex Component 4, DNA, Fungal
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