Error-Prone Translesion DNA Synthesis by Escherichia coli DNA Polymerase IV (DinB) on Templates Containing 1,2-dihydro-2-oxoadenine

Table 2

The insertion kinetics of dCTP and dTTP opposite 1,2-dihydro-2-oxoadenine (2-oxodA) by Pol IV.

Substrate	Km; (fmol/ mL /min)	𝑉max (min^-1)	𝑘cat	𝑘cat/Km
dCTP	8.5	217.4	4.4×10^−3	5.2×10^−4
dTTP	25.6	42.9	8.6×10^−4	3.4×10^−5

Primer 3 was labeled at the 5^-terminal by polynucleotide kinase and annealed with appropriate template oligonucleotides. Primer 3/template 2 (50 fmol) in a 10-𝜇l reaction mixture was incubated at 20^∘C for 60 min with purified Pol IV (50 nM) in the presence of dTTP or dCTP at various concentrations (0.02~500 𝜇M), followed by polyacrylamide gel electrophoresis.