Repair of N-methylpurines in the mitochondrial DNA of xeroderma pigmentosum complementation group D cells.
Previous work from our laboratory has shown that mitochondria are able to repair N-methylpurines formed by methylnitrosourea (MNU). However, it is unclear as to whether repair mechanisms that remove this type of lesion in nuclear DNA also remove these adducts in mitochondria. To address this question, we studied repair of MNU-induced N-methylpurines in the mitochondrial DNA from xeroderma pigmentosum complementation group D (XP-D) cells using quantitative Southern blot analysis and 32P-end-labeling techniques. These cells have been reported to be defective in the repair of this type of lesion in their nuclear genome. WI 38 cells were used as normal controls for these studies. Both XP-D fibroblasts and WI 38 cells were exposed to 0.5 mM MNU for 1 h. Following an 8 h repair period, 61% of N-methylpurines were repaired in the mitochondrial genome of XP-D cells and 39% of these lesions were repaired in WI 38 cells. After 24 h, XP-D cells had repaired 77% of the N-methylpurines in their mitochondrial genome, while WI 38 cells had 44% repair of this type of damage. During this same 24 h time period, 81.5% of the N7-methylguanines had been removed from the total cellular DNA of the WI 38 cells compared to only 38.3% repair of this lesion in the XP-D cells. Thus, XP-D cells, though deficient in the repair of N-methylpurines in their nuclear genome, are proficient in the repair of this type of damage in their mitochondria, suggesting that the mechanisms to repair N-methylpurines in the nuclear and mitochondrial genomes of these cells are different.
Department of Structural and Cellular Biology, University of South Alabama, Mobile 36688.
S P
SP
LeDoux
N J
NJ
Patton
L J
LJ
Avery
G L
GL
Wilson
D015139:Blotting, Southern; D002460:Cell Line; D004249:DNA Damage; D004260:DNA Repair; D004272:DNA, Mitochondrial / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005816:Genetic Complementation Test; D006147:Guanine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008745:Methylation; D008770:Methylnitrosourea / Q000378:metabolism* / Q000494:pharmacology*; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D004272:DNA, Mitochondrial; D006147:Guanine; C008450:7-methylguanine; D008770:Methylnitrosourea