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Excision repair of uv damage in mammalian cells evidence for 2 steps in the excision of pyrimidine dimers



Excision repair of uv damage in mammalian cells evidence for 2 steps in the excision of pyrimidine dimers



Biophysical Journal 22(2): 265-280



The incidence of pyrimidine dimer formation and the kinetics of DNA repair in African green monkey kidney CV-1 cells after UV irradiation were studied by measuring survival, T-4 endonuclease V-sensitive sites, the fraction of pyrimidine dimers in acid-insoluble DNA as determined by TLC and repair replication. CV-1 cells exhibited a survival curve with an extrapolation number of 7.8 and Do [mean lethal dose] of 2.5 J/m2. Pyrimidine dimers were lost from acid-insoluble DNA more slowly than endonuclease-sensitive sites were lost from, or new bases were incorporated into, high MW DNA during the course of repair. Growth of CV-1 cultures in [3H]thymidine or X-irradiation (2 or 10 krad) 24 h before UV irradiation had no effect on repair replication induced by 25 J/m2 of UV. Pyrimidine dimer excision measurements by TLC were probably unaffected by radiation from high levels of incorporated radionuclides. The endonuclease-sensitive site and TLC measurements were reconciled by the assumption that pyrimidine dimers were excised from high MW DNA in acid-insoluble oligonucleotides that were slowly degraded to acid-soluble fragments.

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Accession: 005416764

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