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• W1969030130 endingPage "400" @default.
• W1969030130 startingPage "389" @default.
• W1969030130 abstract "A series of six oligonucleotides with dihydrodiol epoxide metabolites of the polycyclic aromatic hydrocarbons (PAHs) benz[a]anthracene and benzo[a]pyrene attached to adenine N6 and guanine N2 atoms were prepared and studied with the processive bacteriophage DNA polymerase T7, exonuclease- (T7-). HIV-1 reverse transcriptase was much less efficient in polymerization than T7-. Benz[a]anthracene and benzo[a]pyrene adducts strongly blocked incorporation of dTTP and dCTP opposite the A and G derivatives, respectively. dATP was preferentially incorporated in all cases. Steady state kinetic analysis indicated that the low catalytic efficiency with adducted DNA was due to both increased Km and lowered kcat values. Some differences due to PAH stereochemistry were observed. Fluorescence estimates of Kd and presteady state kinetic measurements of koff showed no major decrease in the affinity of T7- with damaged DNA substrates or with dNTPs. Presteady state kinetics showed a lack of the normal burst kinetics for dNTP incorporation with all PAH−DNA derivatives. These results indicate that the rate-limiting step is at or before the step of phosphodiester bond formation; release of the oligonucleotide is no longer the slowest step. Thio elemental effects (substitution of α-oxygen with sulfur) were relatively small, in contrast to previous work with T7- and 8-oxo-7,8-dihydroguanine. The effect of these bulky PAH adducts is either to attenuate rates of conformational changes or to introduce an additional conformation problem but not to alter the inherent affinity of the polymerase for DNA or dNTPs." @default.
• W1969030130 created "2016-06-24" @default.
• W1969030130 creator A5019444857 @default.
• W1969030130 creator A5025977173 @default.
• W1969030130 creator A5059327438 @default.
• W1969030130 date "2005-02-01" @default.
• W1969030130 modified "2023-10-18" @default.
• W1969030130 title "Kinetics of Nucleotide Incorporation Opposite Polycyclic Aromatic Hydrocarbon−DNA Adducts by Processive Bacteriophage T7 DNA Polymerase" @default.
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• W1969030130 doi "https://doi.org/10.1021/tx049683c" @default.
• W1969030130 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/15720147" @default.
• W1969030130 hasPublicationYear "2005" @default.
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