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The preference for error-free or error-prone postreplication repair in Saccharomyces cerevisiae exposed to low-dose

Dongqing Huang1, Brian D Piening, Amanda G Paulovich

  • 1Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.

Molecular and Cellular Biology
|February 6, 2013
PubMed

Insights

Low-dose methyl methanesulfonate (MMS) reveals distinct DNA repair pathways. Postreplication repair (PRR) mutants show sensitivity, with error-free repair favored in S phase and error-prone repair in G2, impacting cell survival and mutation rates.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Cells utilize postreplication repair (PRR) to tolerate DNA damage, involving both error-free and error-prone mechanisms.
  • Low-dose methyl methanesulfonate (MMS) is a unique tool for studying PRR as it specifically impacts PRR mutants without affecting wild-type cells.

Purpose of the Study:

  • To investigate the genome-wide sensitivity of cells to low-dose MMS.
  • To elucidate the distinct roles and preferences of PRR pathways in response to DNA damage at different cell cycle stages.

Main Methods:

  • Genome-wide screening for sensitivity to 0.001% methyl methanesulfonate (MMS).
  • Analysis of PRR-defective mutants (rad18Δ) under low-dose MMS exposure.
  • Investigation of cell cycle arrest, single-stranded DNA gap accumulation, and the roles of Rad18, MEC1, and REV3.

Main Results:

  • PRR-defective mutants exhibit exquisite sensitivity to low-dose MMS, leading to prolonged G2 arrest and single-stranded DNA gap accumulation.
  • Rad18 reactivation restores viability and genome integrity after prolonged G2 arrest.
  • PRR pathway choice is context-dependent: error-free repair is preferred in S phase (MEC1-independent), while error-prone translesion synthesis (REV3-dependent) is utilized in G2, requiring MEC1 and causing hypermutability.

Conclusions:

  • Low-dose MMS is a sensitive probe for dissecting PRR pathway dynamics.
  • The timing of DNA damage and repair influences pathway preference, impacting cellular survival and mutagenic outcomes.
  • Understanding PRR pathway regulation is crucial for comprehending genome stability and mutation processes.

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