Base substitutions, targeted single-base deletions, and chromosomal translocations induced by bleomycin in plateau-phase mammary epithelial cells.
Previous work showed that treatment of plateau-phase Chinese hamster ovary cells with the radiomimetic double-strand cleaving agent bleomycin induced very small deletions as well as interchromosomal reciprocal translocations, both of which could be ascribed to errors in end joining of DNA double-strand breaks. In an attempt to assess the possible role of TP53 in suppressing such repair errors, bleomycin-induced mutagenesis at the HPRT locus was examined in immortalized 184B5 human mammary epithelial cells (TP53(+)), and in a TP53-defective derivative, 184B5-E6tfxc6. For both cell lines, the most frequent bleomycin-induced mutations were base substitutions, with no apparent targeting to major bleomycin lesions. However, both lines also sustained single-base deletions that were targeted to expected sites of double-strand breaks, suggesting that they arose by end-joining repair of the breaks. Surprisingly, only a few large deletions or rearrangements, and no interchromosomal events involving the HPRT locus were detected among the mutants. The results suggest that in both cell lines, errors in double-strand break repair resulting in heritable large deletions and rearrangements are rare. Spectral karyotyping of bleomycin-treated 184B5 cells showed that a significant number of translocations were present shortly after bleomycin exposure, but their frequency decreased upon continued culture of the cells. Thus, for these cells, the lack of induced interchromosomal rearrangements can be explained in part by selection against such events as the cells proliferate.
Yu
Yin
Y
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, Virginia 23298, USA.
Inamdar
Kedar V
KV
Turner
Kristi
K
Jackson-Cook
Colleen K
CK
Povirk
Lawrence F
LF
D001483:Base Sequence; D001761:Bleomycin / Q000494:pharmacology / Q000633:toxicity*; D001940:Breast / Q000166:cytology*; D002461:Cell Line, Transformed / Q000187:drug effects; D004252:DNA Mutational Analysis; D004260:DNA Repair; D004847:Epithelial Cells / Q000187:drug effects; D016193:G1 Phase; D005796:Genes / Q000187:drug effects; D016158:Genes, p53; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D007621:Karyotyping; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000187:drug effects*; D017354:Point Mutation; D012641:Selection, Genetic; D017384:Sequence Deletion; D014178:Translocation, Genetic; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency / Q000502:physiology*
D016159:Tumor Suppressor Protein p53; D001761:Bleomycin; D007041:Hypoxanthine Phosphoribosyltransferase