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Synchronization of S Phase in Schizosaccharomyces pombe Cells by Transient Exposure to M-Factor Pheromone

Olaf Nielsen1

  • 1Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.

Cold Spring Harbor Protocols
|September 3, 2016
PubMed

Insights

Synchronizing DNA replication in fission yeast is challenging. Combining nitrogen starvation with M-factor pheromone treatment effectively synchronizes passage through S phase, aiding DNA replication studies.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Fission yeast offers a model for DNA replication studies due to its cell cycle characteristics and genetic tractability.
  • Synchronizing the cell cycle, particularly S phase, in fission yeast has been a significant experimental hurdle.
  • Understanding DNA replication is crucial for cell division and preventing genetic abnormalities.

Purpose of the Study:

  • To develop a reliable method for synchronizing fission yeast through S phase.
  • To facilitate detailed studies of DNA replication processes in a model organism.
  • To overcome limitations in existing cell cycle synchronization techniques.

Main Methods:

  • Utilizing nitrogen starvation to arrest cells in G1 phase.
  • Employing M-factor mating pheromone to further enhance G1 arrest.
  • Combining both treatments to achieve high-synchrony S phase entry.

Main Results:

  • The combined nitrogen starvation and M-factor treatment effectively synchronizes fission yeast.
  • Cells treated with this protocol exhibit a synchronized passage through an ostensibly normal S phase.
  • This method provides a robust tool for studying DNA replication dynamics.

Conclusions:

  • The described protocol offers a highly effective solution for fission yeast cell cycle synchronization.
  • This advancement enables more precise investigations into DNA replication mechanisms.
  • The method is valuable for researchers studying cell division and genome stability.

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