Mutational specificity of 2-amino-3-methylimidazo-[4,5-f]quinoline in the hprt locus of CHO-K1 cells.

2-Amino-3-methylimidazo[4,5-f]quinoline (IQ), a food carcinogen formed in cooked meats, can induce gene mutation at the hprt locus of CHO-K1 cells in the presence of hepatic S9 mix. To elucidate the molecular nature of IQ-induced mutation, we characterized the entire coding region of the hypoxanthine phosphoribosyl-transferase gene of 23 independent mutants derived from IQ-treated CHO cells by direct sequencing of polymerase chain reaction-amplified cDNA. Ten of the 23 IQ-induced mutants examined contained single base substitutions; one mutant had three single-base substitutions. Among the base substitutions, G.C-->C.G (six of 13) and A.T-->C.G (three of 13) transversions predominated. Most of the base-substitution mutations occurred preferentially at a middle G and had a dA in their 3' ends. Of the 13 other mutations (56.5%), 12 missing one or more complete exons were splice-site mutations, and one mutant had a partial deletion of an exon. A high frequency of complete exon deletion (11 of 12) in exons 2-5 was observed. Interestingly, 75% of the mutants (nine of 12) with splice-site mutations were induced by IQ only at higher concentrations (300-500 microM). This was probably due to the occurrence of GC base-substitution mutations that affected hprt mRNA splicing, especially at the intron-exon boundaries.