Cellular responses and repair of single-strand breaks introduced by UV damage endonuclease in mammalian cells.
Although single-strand breaks (SSBs) occur frequently, the cellular responses and repair of SSB are not well understood. To address this, we established mammalian cell lines expressing Neurospora crassa UV damage endonuclease (UVDE), which introduces a SSB with a 3'-OH immediately 5' to UV-induced cyclobutane pyrimidine dimers or 6-4 photoproducts and initiates an alternative excision repair process. Xeroderma pigmentosum group A cells expressing UVDE show UV resistance of almost the wild-type level. In these cells SSBs are produced upon UV irradiation and then efficiently repaired. The repair patch size is about seven nucleotides, and repair synthesis is decreased to 30% by aphidicolin, suggesting the involvement of a DNA polymerase delta/epsilon-dependent long-patch repair. Immediately after UV irradiation, cellular proteins are poly(ADP-ribosyl)ated. The UV resistance of the cells is decreased in the presence of 3-aminobenzamide, an inhibitor of poly(ADP-ribose) polymerase. Expression of UVDE in XRCC1-defective EM9, a Chinese hamster ovary cell line, greatly sensitizes the host cells to UV, and addition of 3-aminobenzamide results in almost no further sensitization of the cells to UV. Thus, we show that XRCC1 and PARP are involved in the same pathway for the repair of SSBs.
Department of Molecular Genetics, Institute of Development, Aging, and Cancer, Tohoku University, Sendai, 980-8575 Japan.
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Okano
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Kanno
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Nakajima
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Yasui
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D016466:CHO Cells; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis* / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007700:Kinetics; D008322:Mammals; D011994:Recombinant Proteins / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum
D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; D016590:Aphidicolin; D004247:DNA; D004706:Endodeoxyribonucleases; C105073:uve1 protein, S pombe; D006918:Hydroxyurea