Fapy·dG instructs Klenow Exo^- to misincorporate deoxyadenosine

The effects of Fapy·dG (N-(2-deoxy-α,β-d-erythropentofuranosyl)-N-(2,6-diamino-4-hydroxy-5-formamidopyrimidine)) on the activity of Klenow exo- have been determined by using oligonucleotide substrates containing the lesion at a defined site. Fapy·dG inhibits primer polymerization at two positions: nucleotide incorporation opposite the lesion and extension one nucleotide past the lesion. Klenow exo- is inhibited less by Fapy·dG than by its analogue, MeFapy·dG. Fapy·dG instructs the polymerase to misincorporate deoxyadenosine opposite itself 20 times more frequently than does dG. Extension of the primer containing the Fapy·dG:dA base pair is only slightly less efficient than when dC is opposite the lesion. Overall, Fapy·dG increases the probability that Klenow exo- will make a mistake during replication approximately 80-million fold compared to a template containing the native nucleotide, dG.