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Nucleic Acids Research, 2002, Vol. 30, No. 10 2172-2182
© 2002 Oxford University Press

Mammalian Rad51C contributes to DNA cross-link resistance, sister chromatid cohesion and genomic stability

Barbara C. Godthelp1, Wouter W. Wiegant1, Annemarie van Duijn-Goedhart1, Orlando D. Schärer2, Paul P. W. van Buul1, Roland Kanaar2,3 and Malgorzata Z. Zdzienicka1,4,*

1Department of Radiation Genetics and Chemical Mutagenesis, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL, Leiden, The Netherlands, 2Department of Cell Biology and Genetics, Erasmus University, PO Box 1738, 3000 DR Rotterdam, The Netherlands, 3Department of Radiation Oncology, University Hospital Rotterdam/Daniel, The Netherlands and 4Department of Molecular Cell Genetics, The Ludwik Rydygier University of Medical Sciences, Bydgoszcz, Poland

The eukaryotic Rad51 protein is a structural and functional homolog of Escherichia coli RecA with a role in DNA repair and genetic recombination. Five paralogs of Rad51 have been identified in vertebrates, Rad51B, Rad51C, Rad51D, Xrcc2 and Xrcc3, which are also implicated in recombination and genome stability. Here, we identify a mammalian cell mutant in Rad51C. We show that the Chinese hamster cell mutant, CL-V4B, has a defect in Rad51C. Sequencing of the hamster Rad51C cDNA revealed a 132 bp deletion corresponding to an alternatively spliced transcript with lack of exon 5. CL-V4B was hypersensitive to the interstrand cross-linking agents mitomycin C (MMC) and cisplatinum, the alkylating agent methyl methanesulfonate and the topoisomerase I inhibitor campthotecin and showed impaired Rad51 foci formation in response to DNA damage. The defect in Rad51C also resulted in an increase of spontaneous and MMC-induced chromosomal aberrations as well as a lack of induction of sister chromatid exchanges. However, centrosome formation was not affected. Intriguingly, a reduced level of sister chromatid cohesion was found in CL-V4B cells. These results reveal a role for Rad51C that is unique among the Rad51 paralogs.

* To whom correspondence should be addressed at: Department of Radiation Genetics and Chemical Mutagenesis, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL, Leiden, The Netherlands. Tel: +31 71 5276175; Fax: +31 71 5276173; Email: m.z.zdzienicka{at}lumc.nlPresent address:Orlando D. Schärer, Institute of Medical Radiobiology, University of Zürich, August forel Strasse 7, 8008 Zürich, Switzerland


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