Molecular and structural characterization of oxidized ribonucleotide insertion into DNA by human DNA polymerase (cid:2)

During oxidative stress, inflammation, or environmental exposure, ribo- and deoxyribonucleotides are oxidatively mod-ified. 8-Oxo-7,8-dihydro-2 (cid:2) -guanosine (8-oxo-G) is a common oxidized nucleobase whose deoxyribonucleotide form, 8-oxo-dGTP, has been widely studied and demonstrated to be a mutagenic substrate for DNA polymerases. Guanine ribonucleotides are analogously oxidized to r8-oxo-GTP, which can constitute up to 5% of the rGTP pool. Because ribonucleotides are com-monly misinserted into DNA, and 8-oxo-G causes replication errors, we were motivated to investigate how the oxidized ribonucleotide is utilized by DNA polymerases. To do this, here we employed human DNA polymerase (cid:2) (pol (cid:2) ) and characterized r8-oxo-GTP insertion with DNA substrates containing either a templating cytosine (nonmutagenic) or adenine (mutagenic). Our results show that pol (cid:2) has a diminished catalytic efficiency for r8-oxo-GTP compared with canonical deoxyribonucleotides

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