THE MUTATIONAL SPECIFICITY OF 2-(2-FURYL)-3-(5-NITRO-2-FURYL)-ACRYLAMIDE (AF2) IN THE LACI GENE OF ESCHERICHIA-COLI

We have determined the mutational specificity of the 5-nitrofuran derivative furylfuramide (AF2) in the lacI gene of Escherichia coli. Treatment of a DELTA-uvrB, pKM101 strain with 1 M AF2 yielded a mutation frequency approximately 300 times greater than that of untreated controls. Of the 165 AF2-induced mutants analysed by DNA sequencing, 145 were base substitution mutations, 11 were frameshifts, and the base substitution mutations, 11 were frameshifts, and the remainder small deletions, tandem base substitutions and complex mutations. Base substitution occurred primarily (> 93%) at G:C base pairs. The proportions of the various mutations are very similar to those that have been reported for AP sites. We suggest that the principal mechanism for AF2 mutagenesis is the formation of an adduct which depurinates to yield AP sites that serve as a substrate for error-prone repair. Seventy-two of the mutations occurred at four 5'-TGC-3' sites. The majority (10/11) of the frameshift mutations occurred at one such hotspot and could have been templated by an inverted repeat > 100 bp removed from the site of the mutation.