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A key role for replication factor C in DNA replication checkpoint function in fission yeast

N Reynolds1, P A Fantes, S A MacNeill

  • 1Institute of Cell and Molecular Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.

Nucleic Acids Research
|December 24, 1998
PubMed
Summary

Replication Factor C (RFC) subunit 2 is essential for cell viability in fission yeast. Its absence inactivates the DNA replication checkpoint, allowing mitosis with incompletely replicated DNA.

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Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • DNA Replication

Background:

  • Replication factor C (RFC) is a crucial accessory factor for DNA polymerases delta and epsilon.
  • RFC facilitates the loading of the processivity factor PCNA onto nascent DNA strands at the replication fork.

Purpose of the Study:

  • To genetically analyze the rfc2+ gene in Schizosaccharomyces pombe.
  • To investigate the role of RFC subunit 2 in DNA replication and checkpoint control.

Main Methods:

  • Gene deletion analysis of rfc2+ in fission yeast.
  • Observation of cell cycle progression and DNA replication status in rfc2 Delta mutants.

Main Results:

  • Deletion of rfc2+ is lethal in fission yeast.

Related Experiment Videos

  • rfc2 Delta mutants enter mitosis with incompletely replicated DNA.
  • The DNA replication checkpoint is inactive in rfc2 Delta cells.

    • Conclusions:

    • RFC complex assembly on the primer 3'-end is critical for checkpoint activation.
    • RFC subunit 2 plays a vital role in establishing a checkpoint-competent state during DNA replication.