Small deletion and insertion mutations induced by the topoisomerase II inhibitor teniposide in CHO cells and comparison with sites of drug-stimulated DNA cleavage in vitro.
Sixty-five teniposide-induced mutations at the hemizygous aprt locus of CHO D422 cells were analyzed by polymerase chain reaction and DNA sequencing. Most (63%) of the mutations were deletions, duplications and insertions of various sizes, with the majority being less than 20 base-pairs. The remaining mutations were base substitutions, the majority of which were transversions. A significant correspondence was found between the teniposide-induced small deletion/duplication mutations and sites of teniposide-stimulated DNA strand cleavage by topoisomerase II in vitro. In particular, sequences which were deleted in one or more of the mutants showed a much higher incidence of strong cleavage sites than sequences not involved in deletions. However, the exact positioning of the cleavage sites with respect to the deletion termini was variable. The data did not suggest any unified model to account for all the mutations, but most of the deletions and duplications could be accounted for by one of three mechanisms: (1) double-strand break repair nonhomologous end-joining; (2) replication slippage/misalignment; and (3) addition or deletion of a few nucleotides at free 3' ends left by topoisomerase II, as previously suggested for similar mutations in phage T4. There was no evidence that topoisomerase II subunit exchange was a significant mechanism of mutagenesis in this system.
Han
Y H
YH
Department of Pharmacology and Toxicology, Medical College of Virginia Virginia Commonwealth University, Richmond 23928.
Austin
M J
MJ
Pommier
Y
Y
Povirk
L F
LF
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000187:drug effects*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009153:Mutagens / Q000494:pharmacology*; D017384:Sequence Deletion; D013713:Teniposide / Q000494:pharmacology*; D059005:Topoisomerase II Inhibitors
D009153:Mutagens; D059005:Topoisomerase II Inhibitors; D004247:DNA; D013713:Teniposide