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1,N(2)-Etheno(epsilon)guanine (epsilon) is formed in DNA as a result of exposure to certain vinyl monomers (e.g., vinyl chloride) or from lipid peroxidation. This lesion has been shown to be mutagenic in bacteria and mammalian cells. 1,N(2)-epsilon-G has been shown to block several model replicative DNA polymerases (pols), with limited bypass. Recently, an archebacterial DNA pol, Sulfolobus solfataricus Dpo4, has been shown to copy past 1,N(2)-epsilon-G. In this study, we examined the abilities of recombinant, full-length human pol delta and three human translesion DNA pols to copy past 1,N(2)-epsilon-G. The replicative pol, pol delta, was completely blocked. Pols iota and kappa showed similar rates of incorporation of dTTP and dCTP. Pol eta was clearly the most active of these pols in copying past 1,N(2)-epsilon-G, incorporating in the order dGTP &gt; dATP &gt; dCTP, regardless of whether the base 5' of 1,N(2)-epsilon-G in the template was C or T. Pol eta also had the highest error frequency opposite 1,N(2)-epsilon-G. Analysis of the extended products of the pol eta reactions by mass spectrometry indicated only two products, both of which had G incorporated opposite 1,N(2)-epsilon-G and all other base pairing being normal (i.e., G:C and A:T). One-half of the products contained an additional A at the 3'-end, presumably arising from a noninformational blunt end addition or possibly a slipped insertion mechanism at the end of the primer-template replication process. In summary, the most efficient of the four human DNA pols was pol eta, which appeared to insert G opposite 1,N(2)-epsilon-G and then copy correctly. This pattern differs with the same oligonucleotide sequences and 1,N(2)-epsilon-G observed using Dpo4, emphasizing the importance of pols in mutagenesis events.

Translesion Synthesis Across 1,N2-Ethenoguanine by Human DNA Polymerases

Translesion Synthesis Across 1,N²-Ethenoguanine by Human DNA Polymerases