Kinetics of repair of UV-induced DNA damage in repair-proficient and -deficient cells as determined by quantitative polymerase chain reaction.
Advances in methodologies to monitor gene-specific repair in human cells have facilitated a detailed understanding of the complexity of the nucleotide excision repair system. One of these procedures, quantitative polymerase chain reaction (QPCR), holds significant promise for dissecting the fine structure of the repair of UV-induced DNA damage. This assay was used to study the repair of UV photoproducts in both actively transcribed and nontranscribed genes from human cells that were capable of (1) repair of both cyclobutane pyrimidine dimers and 6-4 photoproducts; (2) removal of neither dimers nor 6-4 photoproducts; (3) strong preferential repair of 6-4 photoproducts relative to dimers; and (4) severely depressed rates of 6-4 photoproducts and dimers. Detailed kinetic analyses revealed that repair of both active and inactive genes can be studied with a very fine degree of precision and that the repair status of the cells can easily be detected by use of the procedures described.
McCarthy
M J
MJ
Department of Biochemistry, Vanderbilt University, Nashville, TN, USA.
Rosenblatt
J I
JI
Lloyd
R S
RS
D002453:Cell Cycle; D002460:Cell Line; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D014158:Transcription, Genetic; D014466:Ultraviolet Rays / Q000009:adverse effects*
D004247:DNA