Analysis of mutation spectra in UVB‐exposed mouse skin epidermis and dermis: Frequent occurrence of C→T transition at methylated CpG‐associated dipyrimidine sites

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Abstract
We recently reported the kinetics of mutation induction by UVB in the skin epidermis and dermis of transgenic Muta™ mice [Ikehata and Ono, Mutat Res 508:41–47, 2002]. In the present study we determined the complete DNA sequence of the lacZ transgene in 208 mutants isolated from the dermis and epidermis of UVB‐irradiated and control mice. The resulting mutation patterns for the dermis and epidermis were similar, although two CC→TT tandem substitutions, one of the signature mutations for UV insult, were detected only among the UVB‐induced epidermal mutants. The spectra of the UVB‐induced and control mutations were both dominated by C→T transitions (83% and 62%); however, the C→T transitions from irradiated mice occurred almost exclusively in dipyrimidine sites, while those from control mice preferred CpG sites. Thus, the mutation spectrum detected for the irradiated skin tissues was different from the background spectrum and UV‐specific, confirming the utility of the transgenic system for UVB‐induced mutation studies in vivo. An analysis of the bases adjacent to the mutated cytosines from irradiated mice revealed that the dipyrimidine sites preferred for UVB‐induced mutation were 5′‐TC‐3′ > 5′‐CC‐3′ > 5′‐CT‐3′. Among mutants from irradiated mice, C→T transitions were recovered frequently at dipyrimidine sites associated with CpG. We showed that CpG sites in the lacZ transgene of Muta™ mice were heavily methylated in both the epidermis and dermis. Thus, CpG methylation could contribute to the UVB‐induced recurrent or hotspot mutations in the mammalian genome. Environ. Mol. Mutagen. 41:280–292, 2003.