Formation of quadruplex DNA at a hot-spot for H 2O 2/Fe-mediated mutagenesis

Abstract

A shuttle vector, pZ189, carrying a bacterial suppressor tRNA marker gene (supF) was dissolved in Tris-EDTA buffer containing 0.3 M 10B-enriched boric acid and then irradiated with boron neutron captured beam (BNCB) produced by the nuclear reaction 10B (n,α) 7Li with thermal neutrons. A DNA repair-deficient mutant, KS46 (uvrA−), of Escherichia coli was transformed with the plasmid DNA, and the transformants carrying mutations on the supF Gene were selected as nalidixic acid-resistant colonies. The mutation frequency (2.4 × 10−4) of pZ189 at the D10 dose was about 70 times greater than the spontaneous rate (3.5 × 10−6). The plasmid mutations were analyzed using DNA sequencers; 88% of them were base substitutions. A few minus-one frameshifts (7%) and deletions (5%) were detected. Among these base substitutions, transversions of G:C to T:A (42%) and G:C to C:G (29%) predominated. Twenty-seven percent of the base substitutions were G:C to A:T transitions; no A:T to G:C transitions were detected.