Characterization of mutations induced by 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine in the colon of gpt delta transgenic mouse:: novel G:C deletions beside runs of identical bases

Mutations induced by one of the typical dietary mutagens/carcinogens, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), were characterized using gpt delta transgenic mice. This transgenic mouse model has two selection methods to efficiently detect different types of mutations, i,e, 6-thioguanine selection for point mutations and Sp(-) selection for deletions. The mice were fed with a diet containing 400 p,p,m, PhIP for 13 weeks and gpt and Spi(-) mutations were analyzed from the colon, where the highest mutant frequencies were detected. Concerning the types of gpt mutations from PhIP-treated mice, 81% were single base pair substitutions and G:C --> T:A transversions predominated; single base pair deletions at G:C base pairs were also observed. In untreated mice G:C --> A:T transitions predominated and >80% of these events involved 5'-CpG-3' sites. Concerning Spi(-) mutants from PhIP-treated mice, 76% were G:C base pair deletions and more than half of these events occurred in monotonic G or C run sequences. Interestingly, a novel type of frameshift motif, i.e, G:C base pair deletions beside run sequences, was observed. The most frequently observed mutation in this class was the 5'-TTTTTTG-3' --. 5'-TTTTTT-3' event. These results suggest that PhIP induces point mutations, such as base substitutions and single base pair deletions, rather than larger deletions irt vivo and that run sequences may play an important role in PhIP-induced G:C base pair deletions.