Table 2.

Fidelity of in vitro DNA synthesis by wild-type Polδ and Polδ-R696W

WT Polδ (100 μM dNTPs)Polδ-R696W (100 μM dNTPs)WT Polδ (S-phase dNTPs)Polδ-R696W (S-phase dNTPs)
No.ER (×10−5)No.ER (×10−5)No.ER (×10−5)No.ER (×10−5)
Base substitutions (mispair)321.4775.1341.311815
 Transitions251.5393.5221.27913
  A→G (A-dCTP)0<0.2431.110.2110.67
  G→A (G-dTTP)30.9241.892.54539
  C→T (C-dATP)184.0206.881.6159.4
  T→C (T-dGTP)40.82123.740.731810
 Transversions70.26382.2120.40394.1
  A→C (A-dGTP)0<0.3421.020.6010.94
  A→T (A-dATP30.67124.120.4063.8
  C→A (C-dTTP)0<0.2941.80<0.2697.4
  C→G (C-dCTP)0<0.450<0.680<0.400<1.3
  G→C (G-dGTP)30.8441.751.253.9
  G→T (G-dATP)10.27104.130.721310
  T→A (T-dTTP)0<0.4031.80<0.3533.3
  T→G (T-dCTP)0<0.2731.20<0.2421.5
Frameshifts150.50412.180.24383.5
 Minus 1150.50402.060.18383.5
 Plus 10<0.030<0.0520.060<0.09
Large deletions1013
 Total4811843159
lacZ mutant frequency0.00190.00710.00150.017
  • The S-phase dNTP concentrations used for wild-type Polδ and Polδ-R696W are the same as calculated for wild-type and pol3-R696W + pPOL3 yeast strains (Table 3). All lacZ mutants recovered from reactions with wild-type Polδ contained single mutations. Sixteen percent of mutants obtained from reactions with Polδ-R696W contained more than one mutation. Only phenotypically detectable mutations are listed. Error rates (ERs) for individual mutation types were calculated as described in Materials and Methods. Silent mutations were excluded from the error rate calculation. The location of mutations in the lacZ sequence is shown in Fig. S2, along with the description of large deletions and other rare mutations. The background mutation frequency for unfilled M13mp2 gapped substrate was 0.0016.