Singlet Oxygen Induces Predominantly G to T Transversions on a Single-Stranded Shuttle Vector Replicated in Monkey Cells

Abstract

To elucidate the mechanisms of mutagenesis by singlet oxygen DNA damage in mammalian cells, a SV40-derived single-stranded shuttle vector was exposed to the water soluble endoperoxide 3,3′-(1,4-naphthylidene) dipropionate (NDPO₂). The damaged vector was transfected into monkey COS7 cells and the plasmid progeny exhibited up to 10 fold increase on the mutation frequency in the supF target gene, when compared to untreated vector. The sequence in the supF locus of such mutants revealed that singlet oxygen-induced mutagenesis in single-stranded vector is significantly different from spontaneous mutagenesis. Among the base substitutions, most of the mutations involved deoxyguanosines, being G to T transversions the predominant type of change. The data indicate that mutagenesis by singlet oxygen in mammalian cells may be generated by an error prone bypass of damaged deoxyguanosines at the template DNA.

Key Words:

• Singlet oxygen
• mutagenesis
• DNA damage
• 8-hydroxydeoxyguanine
• shuttle vector