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Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
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The human pre-replication complex is an open complex.

Jian Li1, Jiangqing Dong2, Weitao Wang3

  • 1School of Biological Sciences, The University of Hong Kong, Hong Kong, China.

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|January 7, 2023
PubMed
Summary
This summary is machine-generated.

Researchers uncovered how the minichromosome maintenance (MCM) double hexamer (DH) initiates DNA replication by melting DNA strands. This structure reveals an initial open structure (IOS) crucial for replication licensing in human cells.

Keywords:
DNA replication initiationhuman MCM2–7 complexinitial DNA meltingorigin firingpre-RCreplication licensing

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA replication initiation is essential for cell division in eukaryotes.
  • The minichromosome maintenance (MCM) 2-7 double hexamer (DH) is a key complex for initiating DNA replication.
  • The precise mechanism by which the MCM-DH melts origin DNA remains largely unknown.

Purpose of the Study:

  • To elucidate the structural mechanism of initial DNA melting by the human MCM-DH (hMCM-DH) during replication initiation.
  • To understand how the hMCM-DH facilitates the formation of an initial open structure (IOS) at replication origins.
  • To investigate the role of the IOS in licensing DNA replication.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) to determine the high-resolution structure of the hMCM-DH bound to DNA.
  • Biochemical assays to assess the impact of disturbing the IOS on DH formation and replication initiation.
  • Genomic footprinting to map the distribution of hMCM-DH and IOSs across the genome.

Main Results:

  • A 2.59-Å cryo-EM structure of the hMCM-DH revealed a constricted central channel that distorts bound DNA.
  • The hMCM-DH induces an initial open structure (IOS) by separating one base pair within the distorted DNA duplex.
  • Disruption of the IOS significantly inhibits MCM-DH formation and subsequent replication initiation.
  • Genomic mapping showed IOSs are clustered in initiation zones, correlating with stochastic origin firing.

Conclusions:

  • The hMCM-DH possesses an intrinsic mechanism to couple DNA binding with initial strand separation, forming an IOS.
  • This IOS is critical for licensing DNA replication initiation in human cells.
  • The findings provide a structural basis for understanding how replication origins are selected and opened.