RELATING AROMATIC HYDROCARBON-INDUCED DNA-ADDUCTS AND C-H-RAS MUTATIONS IN MOUSE SKIN PAPILLOMAS - THE ROLE OF APURINIC SITES

Mouse skin tumors contain activated c-H-ras oncogenes, often caused by point mutations at codons 12 and 13 in exon 1 and codons 59 and 61 in exon 2. Mutagenesis by the noncoding apurinic sites can produce G --> T and A --> T transversions by DNA misreplication with more frequent insertion of deoxyadenosine opposite the apurinic site. Papillomas were induced in mouse skin by several aromatic hydrocarbons, and mutations in the c-H-ras gene were determined to elucidate the relationship among DNA adducts, apurinic sites, and ras oncogene mutations. Dibenzo-[a,l]pyrene (DB[a,l]P), DB [a,l]P-11,12-dihydrodiol, anti-DB [a,l]P-11,12-diol-13,14-epoxide, DB [a,l]P-8,9-dihydrodiol, 7,12-dimethylbenz[a]anthracene (DMBA), and 1,2,3,4-tetrahydro-DMBA consistently induced a CAA --> CTA mutation in codon 61 of the c-H-ras oncogene. Benzo[a]pyrene induced a GGC --> GTC mutation in codon 13 in 54% of tumors and a CAA --> CTA mutation in codon 61 in 15%. The pattern of mutations induced by each hydrocarbon correlated with its profile of DNA adducts. For example, both DB[a,l]P and DMBA primarily form DNA adducts at the N-3 and/or N-7 of deoxyadenosine that are lost from the DNA by depurination, generating apurinic sites. Thus, these results support the hypothesis that misreplication of unrepaired apurinic sites generated by loss of hydrocarbon-DNA adducts is responsible for transforming mutations leading to papillomas in mouse skin.