Repair by human cell extracts of single (6-4) and cyclobutane thymine-thymine photoproducts in DNA.
One cis-syn cyclobutane thymine dimer or one (6-4) thymine-thymine photoproduct was built into an identical sequence of a closed-circular M13 duplex DNA, and nucleotide excision repair synthesis carried out by human cell extracts in the area containing each lesion was determined. Extracts from normal cells repaired the (6-4) photoproduct with a patch size of approximately 20-30 nucleotides, but repair was at least 10-fold lower at the cyclobutane dimer. The (6-4) lesion was repaired with comparable efficiency to a single acetylamino-fluorene-guanine adduct in a similar location. Extract from nucleotide excision repair-deficient xeroderma pigmentosum group A cells could not remove any of these adducts but could complete repair of the lesions after incision with Escherichia coli UvrABC proteins. This direct comparison of repair of two UV photoproducts, in an in vitro system where chromatin assembly and transcription are absent, suggests that the more rapid repair of the (6-4) lesion observed in the mammalian cell genome overall is due in part to a significant difference in the ability of the repair complex to locate and incise these lesions in DNA.
Szymkowski
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DE
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, United Kingdom.
Lawrence
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CW
Wood
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RD
D017104:Bacteriophage M13; D001483:Base Sequence; D002460:Cell Line; D004249:DNA Damage; D004260:DNA Repair; D004270:DNA, Circular / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004706:Endodeoxyribonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009838:Oligodeoxyribonucleotides / Q000528:radiation effects; D011740:Pyrimidine Dimers / Q000378:metabolism*; D013941:Thymine; D014466:Ultraviolet Rays
D004270:DNA, Circular; D004279:DNA, Viral; D029968:Escherichia coli Proteins; D009838:Oligodeoxyribonucleotides; D011740:Pyrimidine Dimers; D004706:Endodeoxyribonucleases; C036385:endodeoxyribonuclease uvrABC; D013941:Thymine