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Published on February 29, 2008; 10.1104/pp.108.116806

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Received January 28, 2008
Accepted February 25, 2008

A homologue of ScRAD5 is involved in DNA repair and homologous recombination in Arabidopsis thaliana

I-Peng Chen , Anja Mannuss , Nadiya Orel , Fabian Heitzeberg , and Holger Puchta *

Botanical Institute II, University Karlsruhe, D-76128 Karlsruhe, Germany

* Corresponding author; email: Holger.Puchta{at}botanik2.uni-karlsruhe.de.

RAD5 is the key component in the RAD5-dependent error-free branch of post-replication repair (PRR) in yeast. Rad5 is a member of the Snf2 ATPase/helicase family, possessing as characteristic feature, a RING-finger domain embedded in the SNF2-helicase domain and a HIRAN domain. Yeast mutants are sensitive to DNA damaging agents and reveal differences in homologous recombination. By sequence comparisons we were able to identify two homologues (AtRAD5a and AtRAD5b) in the Arabidopsis genome, sharing about 30% identity and 45% similarity to Saccharomyces cerevisiae Rad5. AtRad5a and AtRad5b have the same kind of domain organization with a higher degree of similarity to each other than to ScRad5. Surprisingly, both genes differ in function: whereas two independent mutants of Atrad5a are hypersensitive to the crosslinking agents mitomycin C and cis-platin and to a lesser extent to the methylating agent, methyl methane sulfonate, the Atrad5b mutants did not exhibit any sensitivity to all DNA damaging agents tested. An Atrad5a/Atrad5b double mutant resembles the sensitivity phenotype of the Atrad5a single mutants. Moreover, in contrast to Atrad5b, the two Atrad5a mutants are deficient in homologous recombination after treatment with the double-strand break inducing agent bleomycin. Our results suggest that the RAD5-dependent error-free branch of PRR is conserved between yeast and plants, and that AtRAD5a might be functionally homologous to ScRAD5.






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