RAD51 is involved in repair of damage associated with DNA replication in mammalian cells.
The RAD51 protein, a eukaryotic homologue of the Escherichia coli RecA protein, plays an important role in the repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) in mammalian cells. Recent findings suggest that HR may be important in repair following replication arrest in mammalian cells. Here, we have investigated the role of RAD51 in the repair of different types of damage induced during DNA replication with etoposide, hydroxyurea or thymidine. We show that etoposide induces DSBs at newly replicated DNA more frequently than gamma-rays, and that these DSBs are different from those induced by hydroxyurea. No DSB was found following treatment with thymidine. Although these compounds appear to induce different DNA lesions during DNA replication, we show that a cell line overexpressing RAD51 is resistant to all of them, indicating that RAD51 is involved in repair of a wide range of DNA lesions during DNA replication. We observe fewer etoposide-induced DSBs in RAD51-overexpressing cells and that HR repair of etoposide-induced DSBs is faster. Finally, we show that induced long-tract HR in the hprt gene is suppressed in RAD51-overexpressing cells, although global HR appears not to be suppressed. This suggests that overexpression of RAD51 prevents long-tract HR occurring during DNA replication. We discuss our results in light of recent models suggested for HR at stalled replication forks.
Lundin
Cecilia
C
Department of Genetic and Cellular Toxicology, Arrhenius Laboratory, Stockholm University, S-106 91 Stockholm, Sweden.
Schultz
Niklas
N
Arnaudeau
Catherine
C
Mohindra
Atul
A
Hansen
Lasse Tengbjerg
LT
Helleday
Thomas
T
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D005047:Etoposide / Q000633:toxicity; D006918:Hydroxyurea / Q000633:toxicity; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D007700:Kinetics; D008957:Models, Genetic; D051135:Rad51 Recombinase; D011995:Recombination, Genetic; D013936:Thymidine / Q000633:toxicity
D004268:DNA-Binding Proteins; D005047:Etoposide; D007041:Hypoxanthine Phosphoribosyltransferase; D051135:Rad51 Recombinase; D013936:Thymidine; D006918:Hydroxyurea