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A mammalian-like DNA damage response of fission yeast to nucleoside analogs

Sarah A Sabatinos1, Tara L Mastro, Marc D Green

  • 1Department of Molecular and Computational Biology, University of Southern California, Los Angeles, CA 90089, USA.

Genetics
|November 15, 2012
PubMed

Insights

Fission yeast, like human cells, shows DNA damage and cell cycle arrest when exposed to nucleoside analogs (BrdU, EdU). This response impacts DNA replication and metabolism research.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Nucleoside analogs are common tools for labeling newly synthesized DNA.
  • Budding yeast typically tolerates these analogs, unlike many other cell types.

Purpose of the Study:

  • To investigate the response of fission yeast (Schizosaccharomyces pombe) to nucleoside analogs 5-bromo-2'-deoxyuridine (BrdU) and 5-ethynyl-2'-deoxyuridine (EdU).
  • To compare fission yeast's response to that of metazoans.

Main Methods:

  • Incorporation of BrdU and EdU into DNA.
  • Assessment of DNA damage response pathways, including Chk1 activation.
  • Analysis of UV light and drug sensitivity.
  • Investigation of replication checkpoint mutants (cds1Δ).
  • Examination of the role of Spd1 (ribonucleotide reductase inhibitor) and thymidine kinase.

Main Results:

  • Nucleoside analog incorporation induced DNA damage and activated the Chk1 DNA damage checkpoint kinase in fission yeast.
  • Cells incorporating analogs showed increased sensitivity to UV light and DNA-damaging agents.
  • The replication checkpoint mutant cds1Δ exhibited an enhanced DNA damage response.
  • BrdU response was modulated by Spd1, indicating dNTP pool imbalance.
  • Excess thymidine induced G1 arrest in wild-type fission yeast expressing thymidine kinase.

Conclusions:

  • Fission yeast exhibits a metazoan-like response to nucleoside analogs, including DNA damage and checkpoint activation.
  • These findings suggest fission yeast is a suitable model for studying DNA replication and damage, similar to mammalian cells.
  • The study highlights implications for understanding dNTP metabolism and the use of nucleoside analogs in research.

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