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Arabidopsis T-DNA insertional lines for CDC25 are hypersensitive to hydroxyurea but not to zeocin or salt stress
Authors:Natasha D Spadafora  John H Doonan  Robert J Herbert  M Beatrice Bitonti  Emily Wallace  Hilary J Rogers  Dennis Francis
Institution:1School of Biosciences, Cardiff University, Main College, Cardiff CF10 3AT, UK;2Department of Cell Biology, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK;3Institute of Science and the Environment, University of Worcester, Henwick Grove, Worcester WR2 6AJ, UK;4Dipartimento di Ecologia, Università della Calabria, Arcavacata di Rende, I-87030 Cosenza, Italy
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.
Keywords:Arabidopsis thaliana  cell-cycle checkpoints  hydroxyurea  root growth  NaCl  zeocin
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