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An explanation for origin unwinding in eukaryotes.

Lance D Langston1,2, Michael E O'Donnell1,2

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Summary
This summary is machine-generated.

The CMG helicase encircles double-stranded DNA and melts it in an Mcm10-dependent manner. This process is crucial for initiating DNA replication forks at origins.

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DNA replicationS. cerevisiaebiochemistrychemical biologyhelicasereplication origin

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Area of Science:

  • Molecular Biology
  • DNA Replication
  • Biochemistry

Background:

  • Eukaryotic DNA replication origins feature head-to-head CMG (Cdc45-MCM-GINS) helicase complexes.
  • Mcm10 protein is a known activator of CMG helicases, enabling them to encircle single-stranded DNA and initiate replication forks.

Purpose of the Study:

  • To investigate the mechanism by which CMG helicases melt double-stranded DNA while encircling it.
  • To determine the role of Mcm10 in the duplex DNA unwinding activity of CMG.

Main Methods:

  • In vitro force-measuring assays using purified *S. cerevisiae* CMG complexes.
  • Analysis of CMG translocation and DNA melting activity on double-stranded DNA.
  • Investigating the effect of Mcm10 on CMG's DNA unwinding capabilities.

Main Results:

  • The *S. cerevisiae* CMG complex tracks with force while encircling double-stranded DNA.
  • In the presence of Mcm10, CMG actively melts long stretches of double-stranded DNA during translocation.
  • CMG predominantly tracks on the 3'-5' strand during duplex DNA translocation.

Conclusions:

  • Head-to-head CMG complexes at replication origins exert forces on opposite DNA strands.
  • Mcm10 is essential for the Mcm10-dependent melting of double-stranded DNA by CMG complexes in a head-to-head orientation.