Figure 2.

DNA substrate specificity of JGT. (A) Recombinant JGT or (B) T4 βGT and UDP-[³H]glucose incubated with 36 nt-long dsDNA substrate containing one 5hmU or 5hmC residue, as described in the “Materials and methods” section. The modified base within the dsDNA substrate was present in the context of a matched base pair (5hmU:A, 5hmC:G) or mismatched base pair (5hmU:G). CPM were measured and converted into micromolar glucose transferred. All experiments were performed in triplicate, error bars represent standard deviations. (C) UDP-Glo assay of glucosyltransferase activity of JGT for dsDNA substrates used above as described in the “Materials and methods” section. The amount of UDP Cleaved, indicative of the transfer of glucose to DNA, was estimated from a standard curve of UDP. All experiments were performed in triplicate and error bars are representative of standard deviation.