DNA sequence dependence of closely opposed cyclobutyl pyrimidine dimers induced by UV radiation.
Treatment of UV-irradiated DNAs with Micrococcus luteus pyrimidine dimer-DNA glycosylase results in the formation of double-strand breaks due to cleavage at closely opposed pyrimidine dimers. To determine if the induction of closely opposed dimers is significantly affected by DNA nucleotide sequence, end-labeled DNA fragments of known nucleotide sequence were UV irradiated, incubated with pyrimidine dimer-DNA glycosylase, and analyzed by electrophoresis through nondenaturing polyacrylamide gels. Distinct bands of increased electrophoretic mobility were observed, indicating that bifilar cleavage had occurred with greater probability at specific sites in each DNA sequence. In vitro enzymatic photoreactivation of dimers prior to treatment with pyrimidine dimer-DNA glycosylase prevented the appearance of bands. DNA sequence analysis revealed the presence of closely opposed runs of pyrimidines at sites of more frequent bifilar cleavage. Our results indicate that the induction of closely opposed dimers occurs with greater probability at specific sites in DNA sequences and that such sites are characterized by the presence of closely opposed pyrimidine runs.
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D004249:DNA Damage; D045647:DNA Glycosylases; D004269:DNA, Bacterial / Q000528:radiation effects*; D004279:DNA, Viral / Q000528:radiation effects*; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D011740:Pyrimidine Dimers / Q000032:analysis* / Q000378:metabolism; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004279:DNA, Viral; D011740:Pyrimidine Dimers; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; C036401:deoxyribopyrimidine endonucleosidase