A global DNA repair mechanism involving the Cockayne syndrome B (CSB) gene product can prevent the in vivo accumulation of endogenous oxidative DNA base damage.
The Cockayne syndrome B (CSB) gene product is involved in the repair of various types of base modifications in actively transcribed DNA sequences. To investigate its significance for the repair of endogenous oxidative DNA damage, homozygous csb(-/-)/ogg1(-/-) double knockout mice were generated. These combine the deficiency of CSB with that of OGG1, a gene coding for the mammalian repair glycosylase that initiates the base excision repair of 7,8-dihydro-8-oxoguanine (8-oxoG). Compared to ogg1(-/-) mice, csb(-/-)/ogg1(-/-) mice were found to accumulate with age severalfold higher levels of oxidited purine modifications in hepatocytes, splenocytes and kidney cells. In contrast, the basal (steady-state) levels of oxidative DNA modifications in cells from csb(-/-) mice were not different from those in wild-type mice and did not increase with age. The analysis of the repair rates of additional oxidative DNA base modifications induced by photosensitization in immortalized embryonic fibroblasts was in accordance with these findings: compared to wild-type cells, the global repair was only slightly affected in csb(-/-) cells, more compromised in ogg1(-/-) cells, but virtually absent in csb(-/-)/ogg1(-/-) cells. An inhibition of transcription by alpha-amanitin did not block the Csb-dependent repair in ogg1(-/-) fibroblasts. The influence of Csb on the global repair of 8-oxoG was not detectable in assays with total protein extracts and in a shuttle vector system. The data indicate a role for Csb in the removal of 8-oxoG from the overall genome that is independent of both Ogg1-mediated base excision repair and regular transcription.
Osterod
Marcel
M
Institute of Pharmacy, University of Mainz, D-55099 Mainz, Germany.
Larsen
Elisabeth
E
Le Page
Florence
F
Hengstler
Jan G
JG
Van Der Horst
Gijsbertus T J
GT
Boiteux
Serge
S
Klungland
Arne
A
Epe
Bernd
B
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D001483:Base Sequence; D004249:DNA Damage; D004265:DNA Helicases / Q000235:genetics*; D017931:DNA Primers; D004260:DNA Repair; D045643:DNA Repair Enzymes; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D006720:Homozygote; D051379:Mice; D018345:Mice, Knockout; D018384:Oxidative Stress; D000075223:Poly-ADP-Ribose Binding Proteins; D014158:Transcription, Genetic
D017931:DNA Primers; D000075223:Poly-ADP-Ribose Binding Proteins; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004265:DNA Helicases; C513075:Ercc6 protein, mouse; D045643:DNA Repair Enzymes; D003849:Deoxyguanosine