Overexpression of human OGG1 in mammalian cells decreases ultraviolet A induced mutagenesis.
7,8-dihydro-8-oxo-guanine (8-oxoG) is a mutagenic DNA lesion that is induced by ultraviolet A (UVA) radiation. 8-oxoG results in increased frequency of GC-->TA transversion mutations. UVA-induced mutant frequency was measured in the guanine phosphoribosyl transferase (gpt) gene of Chinese hamster ovary cells (AS52) that were stably transfected to overexpress the hOGG1 protein, the human DNA repair glycosylase for 8-oxoG. This mutant frequency was compared with ultraviolet A-induced mutant frequency in AS52 cells stably transfected with the same vector without the hOGG1 gene. The mutant frequency was significantly decreased in the hOGG1 overexpressing cells irradiated with 300 and 400 kJ/m2 ultraviolet A radiation, corresponding to 25% and 10% cell survival, respectively. The hOGG1 overexpressing cells repaired oxidative DNA lesions three times faster than the vector only cells as measured by a semi-automated alkaline elution assay using FPG enzyme, the bacterial OGG1 analogue, to cut DNA at oxidative base modifications. Thus, the lower mutation frequency in UVA-induced mutations of the hOGG1 overexpressing cells may be related to the increased repair of 8-oxoG. No GC-->TA mutations were detected in the UVA-irradiated hOGG1 overexpressing cells. The results suggest a link between the 8-oxoG lesion and UVA-induced mutagenesis. We propose that hOGG1 has a role in maintaining genomic stability in mammalian cells after oxidative stress.
Dahle
Jostein
J
Department of Radiation Biology, The Norwegian Radium Hospital, 0310 Oslo, Norway. jostein.dahle@rr-research.no
Brunborg
Gunnar
G
Svendsrud
Debbie H
DH
Stokke
Trond
T
Kvam
Egil
E
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics / Q000528:radiation effects; D006224:Cricetinae; D003412:Cricetulus; D045647:DNA Glycosylases / Q000235:genetics*; D004260:DNA Repair; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D006151:Guanosine / Q000031:analogs & derivatives / Q000235:genetics; D006801:Humans; D016296:Mutagenesis; D018384:Oxidative Stress / Q000187:drug effects; D017422:Sequence Analysis, DNA; D014162:Transfection; D014466:Ultraviolet Rays / Q000009:adverse effects*
D006151:Guanosine; C046215:8-hydroxyguanosine; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human