Periodic changes of chromatin organization associated with rearrangement of repair patches accompany DNA excision repair of mammalian cells.
We have used 8-methoxypsoralen to probe the chromatin structure of mammalian cells in situ while they repair pyrimidine dimers or bulky lesions in DNA. We observed that excision repair of these DNA lesions is accompanied by periodic alterations of chromatin organization. In parallel, fluctuations of the rates of repair patch synthesis accompanied these structural changes. Taking advantage of the accessibility of free DNA domains for psoralen intercalation, we have developed a technique to quantitatively isolate the micrococcal nuclease-sensitive, free DNA fraction of native bulk chromatin. We have determined the location of newly synthesized repair patches relative to free DNA domains as a function of repair time. Extensive rearrangements of repair patches from these domains into micrococcal nuclease-resistant DNA were observed. Our results indicate that periodic changes of chromatin organization associated with rearrangement of repair patches accompany the process of excision repair in mammalian cells.
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D000818:Animals; D002478:Cells, Cultured; D002843:Chromatin / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism; D004260:DNA Repair / Q000528:radiation effects; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008297:Male; D008730:Methoxsalen / Q000494:pharmacology*; D008836:Micrococcal Nuclease; D051381:Rats; D011919:Rats, Inbred Strains; D014466:Ultraviolet Rays
D002843:Chromatin; D004247:DNA; D008836:Micrococcal Nuclease; D008730:Methoxsalen