Repair of mitochondrial DNA after various types of DNA damage in Chinese hamster ovary cells.
Using methodology recently developed to assess gene-specific DNA repair, we have demonstrated that it is possible not only to study mitochondrial DNA repair, but also directly to compare mitochondrial and nuclear DNA repair in the same biological sample. Complex enzymatic mechanisms recognize and repair nuclear DNA damage, but it has long been thought that there was no DNA repair in mitochondria. Therefore, in an attempt to delineate more clearly which DNA repair mechanisms, if any, are functioning in mitochondria, we have investigated the repair of several specific DNA lesions in mitochondrial DNA. They include cyclobutane dimers, cisplatin intrastrand adducts, cisplatin interstrand crosslinks and alkali-labile sites. We find that pyrimidine dimers and complex alkylation damage are not repaired in mitochondrial DNA, and that there is minimal repair of cisplatin intrastrand crosslinks. In contrast, there is efficient repair of cisplatin interstrand crosslinks as evidenced by approximately 70% of the lesions being removed by 24 h. Additionally, there is efficient repair of N-methylpurines following exposure to methylnitrosourea with approximately 70% of the lesions being removed by 24 h. The results of these studies reveal that repair capacity of mitochondrial DNA damage depends upon the type of lesion produced by the damaging agent. We speculate that a process similar to the base excision mechanism for nuclear DNA exists for mitochondrial DNA but that there is no nucleotide excision repair mechanism to remove more bulky lesions in this organelle.
LeDoux
S P
SP
Department of Structural and Cellular Biology, University of South Alabama, Mobile 36688.
Wilson
G L
GL
Beecham
E J
EJ
Stevnsner
T
T
Wassermann
K
K
Bohr
V A
VA
D000818:Animals; D016466:CHO Cells; D002467:Cell Nucleus / Q000378:metabolism; D002945:Cisplatin; D006224:Cricetinae; D004247:DNA; D018736:DNA Adducts; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D004272:DNA, Mitochondrial / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D009153:Mutagens / Q000633:toxicity; D011740:Pyrimidine Dimers; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014466:Ultraviolet Rays
D018736:DNA Adducts; D004272:DNA, Mitochondrial; D009153:Mutagens; D011740:Pyrimidine Dimers; C057270:cisplatin-DNA adduct; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D002945:Cisplatin