Arabidopsis T-DNA insertional lines for CDC25 are hypersensitive to hydroxyurea but not to zeocin or salt stress

Natasha D Spadafora, John H Doonan, Robert J Herbert, M Beatrice Bitonti, Emily Wallace, Hilary J Rogers, Dennis Francis

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

BACKGROUND AND AIMS: In yeasts and animals, cyclin-dependent kinases are key regulators of cell cycle progression and are negatively and positively regulated by WEE1 kinase and CDC25 phosphatase, respectively. In higher plants a full-length orthologue of CDC25 has not been isolated but a shorter gene with homology only to the C-terminal catalytic domain is present. The Arabidopis thaliana;CDC25 can act as a phosphatase in vitro. Since in arabidopsis, WEE1 plays an important role in the DNA damage/DNA replication checkpoints, the role of Arath;CDC25 in conditions that induce these checkpoints or induce abiotic stress was tested. Methods arath;cdc25 T-DNA insertion lines, Arath;CDC25 over-expressing lines and wild type were challenged with hydroxyurea (HU) and zeocin, substances that stall DNA replication and damage DNA, respectively, together with an abiotic stressor, NaCl. A molecular and phenotypic assessment was made of all genotypes Key

RESULTS: There was a null phenotypic response to perturbation of Arath;CDC25 expression under control conditions. However, compared with wild type, the arath;cdc25 T-DNA insertion lines were hypersensitive to HU, whereas the Arath;CDC25 over-expressing lines were relatively insensitive. In particular, the over-expressing lines consistently outgrew the T-DNA insertion lines and wild type when challenged with HU. All genotypes were equally sensitive to zeocin and NaCl.

CONCLUSIONS: Arath;CDC25 plays a role in overcoming stress imposed by HU, an agent know to induce the DNA replication checkpoint in arabidopsis. However, it could not enhance tolerance to either a zeocin treatment, known to induce DNA damage, or salinity stress.

Original languageEnglish
Pages (from-to)1183-1192
Number of pages10
JournalAnnals of Botany
Volume107
Issue number7
Early online date20 Jul 2010
DOIs
Publication statusPublished - 07 May 2011

Keywords

  • Arabidopsis
  • Arabidopsis Proteins
  • Bleomycin
  • DNA, Bacterial
  • Gene Expression Regulation, Plant
  • Genotype
  • Hydroxyurea
  • Multienzyme Complexes
  • Mutagenesis, Insertional
  • Oxidoreductases
  • Phenotype
  • Plant Roots
  • RNA, Messenger
  • Sodium Chloride
  • Stress, Physiological
  • cdc25 Phosphatases

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