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Replication in hydroxyurea: it's a matter of time.

Gina M Alvino1, David Collingwood, John M Murphy

  • 1Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.

Molecular and Cellular Biology
|July 20, 2007
PubMed
Summary

Hydroxyurea (HU) slows DNA replication by affecting elongation and initiation. This study shows HU delays, but does not alter, the temporal order of DNA replication origin activation genome-wide.

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Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • Genomics

Background:

  • Hydroxyurea (HU) is a known inhibitor of DNA replication, impacting both elongation and initiation phases.
  • HU triggers the intra-S phase checkpoint, a cellular mechanism that monitors DNA replication.
  • Prior studies in budding yeast indicated that HU, under short exposure, inhibits initiation at late S phase origins.

Purpose of the Study:

  • To investigate the genome-wide effects of extended hydroxyurea (HU) exposure on DNA replication origin activation.
  • To determine if HU specifically excludes initiation from late S phase origins.
  • To revise the model of the intra-S phase checkpoint response to HU.

Main Methods:

  • Microarray experiments were utilized to monitor the activation status of all replication origins.
  • Analysis focused on the temporal progression of DNA synthesis under extended HU treatment.
  • Budding yeast was used as the model organism.

Main Results:

  • Genome-wide DNA synthesis and origin activation patterns under HU treatment mirrored those without HU, albeit on a delayed timescale.
  • No specific exclusion of initiation from late S phase origins was observed.
  • All temporal classes of replication origins were affected, but their activation sequence remained consistent.

Conclusions:

  • Hydroxyurea (HU) treatment results in a slowed progression of S phase rather than a specific blockage of late origin firing.
  • The temporal program of replication origin activation is maintained, despite significant delays, under extended HU exposure.
  • A revised model is proposed where the intra-S phase checkpoint orchestrates a slowed, but ordered, replication process in response to HU.