Fig. 7

PykA stimulates the DNA polymerase activity of DnaE. A Representative alkaline agarose gels showing DnaE primer extension time courses (30, 60, 90, 120, and 150 s) with DnaE (10 nM) alone and in the presence or absence of PykA (40 nM tetramer) or BSA (40 nM), as described in “Methods”. Lanes M and C represent DNA size markers and the control radioactive substrate in the absence of any proteins, respectively. B Representative alkaline agarose gels (from three independent experiments) showing DnaE primer extension time courses (30, 60, 90, 120, and 150 s) with DnaE (10 nM) and in the presence or absence of PykA (40 nM tetramer), with molar ratios of DnaE monomer: DnaN dimer: HolB monomer:YqeN monomer:DnaX trimer:PykA tetramer set to 1:1:1:1:1:1, considering the oligomeric states of these proteins. C As in B with 2 nM DnaE and molar ratios of 1:1:1:1:1:1