Increased sensitivity to killing by restriction enzymes in the XR-1 DNA double-strand break repair-deficient mutant.
Repair or misrepair of DNA double-strand breaks (DSBs) is critical in determining cellular survival after gamma-irradiation. In this report, we focus on the cellular and biochemical consequences of restriction enzyme induced DSBs in wild-type Chinese hamster ovary (CHO) cells and the DNA DSB repair-defective mutant XR-1. We find that XR-1 possesses reduced cellular survival after the introduction of restriction enzymes that produce either cohesive or blunt ends. XR-1's sensitivity to killing by restriction enzymes strongly mimics its response to gamma-rays. Using pulsed field electrophoresis, we find that for each enzyme, similar numbers of DNA DSBs are being introduced in both cell lines. The simplest explanation for the increased sensitivity to restriction enzymes in the mutant is that the biochemical defect in XR-1 is not confined to the repair of ionizing radiation induced ends, but extends to DSBs that possess ligatable 3'-hydroxyl and 5'-phosphate ends as well.
Giaccia
A J
AJ
Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104.
MacLaren
R A
RA
Denko
N
N
Nicolaou
D
D
Stamato
T D
TD
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002869:Chromosome Aberrations; D004249:DNA Damage; D004260:DNA Repair; D004262:DNA Restriction Enzymes / Q000494:pharmacology*; D015246:Deoxyribonuclease EcoRI / Q000235:genetics* / Q000494:pharmacology; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D006064:Gonadotropins, Equine / Q000235:genetics; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection
D006064:Gonadotropins, Equine; D005954:Glucosephosphate Dehydrogenase; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI