Completion of excision repair patches in human cell preparations: identification of a probable mode of excision and resynthesis.
Excision repair of u.v. damage in human fibroblasts is more sensitive to inhibitors of DNA polymerase alpha (cytosine arabinoside, aphidicolin) than to an inhibitor of polymerase beta (dideoxythymidine), which indicates a greater role in repair for polymerase alpha than for polymerase beta. These inhibitors all generate shortened patches with free 3' termini; the detailed structure of these patches was investigated in permeable cells or isolated nuclei by degradation of DNA with exonuclease III and by resynthesis with DNA polymerase I (Klenow fragment) and T4 DNA ligase. The structure of the shortened patches appears to be a short stretch of DNA synthesized in the 5'----3' direction within a longer single-strand gap. The single-strand gap ahead of the 3' terminus can be bridged only by the combined action of polymerase and ligase. This structure implies that excision must involve removal of an oligonucleotide or widening of a gap by 5'----3' exonuclease action to produce a single-strand region wide enough to be a substrate for polymerase alpha. There is no evidence for structures generated by nick translation or strand displacement.
D016590:Aphidicolin; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000528:radiation effects*; D003561:Cytarabine / Q000633:toxicity*; D004256:DNA Polymerase I / Q000037:antagonists & inhibitors*; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors*; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects*; D015224:Dideoxynucleosides; D004224:Diterpenes / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D013936:Thymidine / Q000031:analogs & derivatives* / Q000633:toxicity; D014466:Ultraviolet Rays
D015224:Dideoxynucleosides; D004224:Diterpenes; D003561:Cytarabine; C019336:2',3'-dideoxythymidine; D016590:Aphidicolin; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D013936:Thymidine