Figure 6.
The IDCL and C‐terminal region of EhPCNA are needed for enzymatic activity stimulation of EhDNAligI. Reaction containing 500 fmol of [γ‐32P]‐labeled DNA substrate, 50 fmol of EhDNAligI (A), ΔPIP (B), or ΔDBD (C) in the presence of 150 fmol of wild‐type EhPCNA, an IDCL‐binding‐deficient mutant (EhPCNA‐V127A‐I129A), and two C‐terminal interacting region (EhPCNA‐Δ254 and EhPCNA‐P253A‐F254A). Reactions were incubated at room temperature for 0, 1, 2, 4, 6, and 8 min, aliquots were quenched at the indicated times adding stop buffer and loaded into a 15% polyacrylamide/8M urea gels. Data are presented as pmol of ligated product as function of incubation time.