Thermostable DNA polymerases can perform translesion synthesis using 8-oxoguanine and tetrahydrofuran-containing DNA templates.
The translesion synthesis (TLS) capacity of the thermostable DNA polymerases Taq, Tte and Tte-seq utilizing a synthetic abasic site, tetrahydrofuran (THF), and an 8-oxoguanine-containing DNA template was investigated. Measurements with human DNA polymerase beta were used as a "positive control". Thermostable DNA polymerases were observed to perform TLS with different specificities on both substrates. With a THF-containing template, dGMP was preferentially inserted by all the DNA polymerases. In the presence of Mn(II) as a cofactor, all the polymerases incorporated dCMP opposite 8-oxoguanine whereas, in the presence of Mg(II) ions, dAMP was incorporated. It was found that none of the thermophilic DNA polymerases utilized dTTP with either an 8-oxoguanine or a THF-containing template. In all cases, DNA duplex containing THF as damage was processed to full length less effectively than DNA duplex containing 8-oxoguanine.
Belousova
Ekaterina A
EA
Institute of Chemical Biology and Fundamental Medicine, Siberian Division of Russian Academy of Sciences, Lavrentieva Prospect 8, Novosibirsk 630090, Russia.
Rechkunova
Nadejda I
NI
Lavrik
Olga I
OI
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D019951:DNA Polymerase beta / Q000378:metabolism; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism; D004795:Enzyme Stability; D005663:Furans / Q000737:chemistry; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D019914:Taq Polymerase / Q000737:chemistry / Q000378:metabolism; D013824:Thermus / Q000201:enzymology; D016963:Thermus thermophilus / Q000201:enzymology
D003854:Deoxyribonucleotides; D005663:Furans; D011994:Recombinant Proteins; C018674:tetrahydrofuran; C024829:8-hydroxyguanine; D006147:Guanine; D004247:DNA; D019914:Taq Polymerase; D019951:DNA Polymerase beta; D004259:DNA-Directed DNA Polymerase