Table 1.
DNA polymerase φ activity is stimulated by yeast PCNA
DNA polymerase | PCNA addition | DNA
synthesis*
|
|
---|---|---|---|
−Aphidicolin, pmol (fold stimulation) | +Aphidicolin, pmol | ||
GST-Pol5p | − | 2.5 (1) | 0.5 |
GST-Pol5p | + | 12.1 (4.8) | 1.3 |
Polφ | − | 7.0 (1) | 0.4 |
Polφ | + | 40.0 (5.7) | 1.3 |
Pol5dn† | − | 0.1 (1) | 0.1 |
Pol5dn | + | 0.1 (1) | 0.12 |
Polδ | − | 27.1 (1) | 1.4 |
Polδ | + | 163.0 (6.0) | 2.1 |
Polɛ | − | 35.2 (1) | 0.8 |
Polɛ | + | 40.2 (1.1) | 0.9 |
DNA synthesis was measured in the presence of poly(dA)500⋅oligo(dT)10 (5:1) as a template-primer.
Because the purity of the Pol5dn polypeptide was not the same as that of Polφ, the concentration of Pol5dn was estimated by Western blot using mouse polyclonal antibodies against GST-Pol5p, and the same amount of Pol5dn as Polφ was used for the DNA polymerase assay.