Fanconi's anaemia cells have normal steady-state levels and repair of oxidative DNA base modifications sensitive to Fpg protein.
Cells from Fanconi's anaemia (FA) patients are abnormally sensitive to oxygen. However, a distinct genetic defect in either the cellular defence against reactive oxygen species (ROS) or in their metabolic generation has not been identified to date. Recently, the gene for the human 8-hydroxyguanine (8-oxoG) glycosylase, which removes this oxidative base modification from the genome, has been localized on chromosome 3p25, i.e., in the same region as the FA complementation group D (FAD) gene. We therefore studied the removal of photosensitization-induced 8-oxoG residues from the DNA of FA cells, using Fpg protein, the bacterial 8-oxoG glycosylase, to quantify the lesions by alkaline elution. Similar repair kinetics (approx. 50% removal within 2 h) were observed in Epstein-Barr virus (EBV) immortalized lymphoid cells from FA complementation groups A, B, C and D and in control cells from normal donors, as well as in primary fetal lung fibroblasts not yet assigned to a specific complementation group. The susceptibility for the induction of oxidative DNA modifications by photosensitization was similar in all cells. In addition, the background (steady-state) levels of Fpg-sensitive oxidative DNA base modifications, which reflect the balance between generation and removal of the lesions, were similar in control and FA cells. It is concluded that both the generation and the overall removal of 8-oxoG residues in nuclear DNA is not impaired in FA cells.
Will
O
O
Institute of Pharmacy, University of Mainz, Germany.
Schindler
D
D
Boiteux
S
S
Epe
B
B
D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D004260:DNA Repair; D043382:DNA-Formamidopyrimidine Glycosylase; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D008027:Light; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D018384:Oxidative Stress; D017319:Photosensitizing Agents / Q000494:pharmacology
D017319:Photosensitizing Agents; C024829:8-hydroxyguanine; D006147:Guanine; D004247:DNA; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase