Comparison of DNA-incising capacities in fibroblast strains from the Mannheim XP collection after treatment with N-acetoxy-2-acetylaminofluorene and UV light.
The DNA-incising capacity was determined in 8 normal and 23 XP fibroblast strains of the Mannheim XP collection using the alkaline elution technique after treatment with both UV light and the "UV-like" carcinogen (Ac)2ONFln. Experimental conditions were chosen to allow for selective monitoring of repair-specific enzyme-catalyzed breaks. In order to compare DNA-incising capacities of the various cell strains after UV irradiation with those after treatment with (Ac)2ONFln, dose-response experiments including up to 8 dose levels were performed. The elution curves were analyzed by linear regression analysis. Elution velocities (in terms of DNA single-strand breaks per 10(6) nucleotides) were plotted against the square root of the doses. The slope of the resulting regression line yielded a characteristic term, designated EO, for the DNA-incising capacity of each cell strain. In contrast to normal fibroblasts, EO was found to be reduced in all XP cell strains belonging to the complementation groups A, C, D, E, F (or G) and I investigated, after treatment with both UV light or (Ac)2ONFln. Surprisingly, XP variant strains also exhibited lower EO values. A comparison of post-UV with post-(Ac)2ONFln DNA-incising capacities revealed that reduction in the EO values was very similar in all XP cell strains tested. These data suggest that the sensitivity of XP cells towards UV light or (Ac)2ONFln is due to the same enzymatic defect, namely impaired incision of DNA containing pyrimidine dimers or (Ac)2ONFln-DNA adducts.
Popanda
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German Cancer Research Centre, Institute of Biochemistry, Heidelberg.
Thielmann
H W
HW
D015073:2-Acetylaminofluorene / Q000031:analogs & derivatives*; D000099:Acetoxyacetylaminofluorene / Q000378:metabolism*; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D000099:Acetoxyacetylaminofluorene; D004247:DNA; D015073:2-Acetylaminofluorene