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An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
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Related Experiment Video

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Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
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Xenopus Mcm10 is a CDK-substrate required for replication fork stability.

Gaganmeet Singh Chadha1, Agnieszka Gambus1, Peter J Gillespie1

  • 1a Centre for Gene Regulation & Expression, School of Life Sciences, University of Dundee , Dundee , UK.

Cell Cycle (Georgetown, Tex.)
|June 22, 2016
PubMed
Summary

Mcm10 protein is not essential for initiating DNA replication but is crucial for maintaining replication fork stability and replisome integrity, especially under stress. Its recruitment to chromatin requires DDK and CDK activity.

Keywords:
CDKDNA replicationMcm10Replication forkXenopus

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Mcm10's role in DNA replication is debated, particularly in metazoans.
  • Mcm10 is implicated in forming and stabilizing the replicative helicase and replisome.

Purpose of the Study:

  • To investigate the role and regulation of Mcm10 in Xenopus egg extracts.
  • To clarify Mcm10's function in DNA replication initiation and elongation.

Main Methods:

  • Chromatin recruitment assays in Xenopus egg extracts.
  • Depletion of Mcm10 using specific extracts.
  • Analysis of replication fork elongation rates.
  • Assessment of replisome protein stability.

Main Results:

  • Xenopus Mcm10 is recruited late to chromatin, dependent on DDK and CDK activity.
  • Mcm10 is a CDK substrate but chromatin association does not require phosphorylation.
  • Mcm10 depletion does not prevent bulk DNA replication but reduces elongation rate.
  • Absence or lack of Mcm10 phosphorylation leads to replisome instability, especially under replication stress.

Conclusions:

  • Mcm10 is not essential for replication initiation but plays a vital role in fork progression and stability.
  • Mcm10 phosphorylation by CDK influences replisome stability under replication stress.