Functional nucleotide excision repair is required for the preferential removal of N-ethylpurines from the transcribed strand of the dihydrofolate reductase gene of Chinese hamster ovary cells.
Transcription-coupled repair of DNA adducts is an essential factor that must be considered when one is elucidating biological endpoints resulting from exposure to genotoxic agents. Alkylating agents comprise one group of chemical compounds which modify DNA by reacting with oxygen and nitrogen atoms in the bases of the double helix. To discern the role of transcription-coupled DNA repair of N-ethylpurines present in discrete genetic domains, Chinese hamster ovary cells were exposed to N-ethyl-N-nitrosourea, and the clearance of the damage from the dihydrofolate reductase gene was investigated. The results indicate that N-ethylpurines were removed from the dihydrofolate reductase gene of nucleotide excision repair-proficient Chinese hamster ovary cells; furthermore, when repair rates in the individual strands were determined, a statistically significant bias in the removal of ethyl-induced, alkali-labile sites was observed, with clearance occurring 30% faster from the transcribed strand than from its nontranscribed counterpart at early times after exposure. In contrast, removal of N-ethylpurines was observed in the dihydrofolate reductase locus in cells that lacked nucleotide excision repair, but both strands were repaired at the same rate, indicating that transcription-coupled clearance of these lesions requires the presence of active nucleotide excision repair.
Sitaram
A
A
Department of Biology, New York University, New York, New York 10003, USA.
Plitas
G
G
Wang
W
W
Scicchitano
D A
DA
D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D018736:DNA Adducts / Q000235:genetics*; D004260:DNA Repair / Q000502:physiology*; D005038:Ethylnitrosourea / Q000494:pharmacology; D005796:Genes / Q000235:genetics; D011687:Purines / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014158:Transcription, Genetic
D000477:Alkylating Agents; D018736:DNA Adducts; D011687:Purines; D013762:Tetrahydrofolate Dehydrogenase; D005038:Ethylnitrosourea