TABLE 4.
Primase-coupled polymerase activity with human pol α
| Primase | Polymerase | DNA template | Primers utilized | Primers Transferred Intramolecularly |
|---|---|---|---|---|
| % | % | |||
| UL5-UL52-UL8 (Hel-) | pol α | T20GTCCT36 | 19 | <0.001a |
| UL5-UL52-UL8 | pol α | T20GTCCT36 | 29 ± 11 | <0.001 |
| UL5-UL52-UL8 | pol α-primase | T20GTCCT36 | 18 ± 5 | -b |
| UL5-UL52-UL8 | UL30-UL42 + pol α | T20GTCCT36 | 10 ± 5 | 0.12 ± 0.06 |
| UL5-UL52-UL8 | UL30-UL42 + pol α-primase | T20GTCCT36 | 8 ± 3 | -b |
| UL5-UL52 | pol α | T20GTCCT36 | 7.9 ± 0.7 | <0.001 |
| UL5-UL52 + UL8 | pol α | T20GTCCT36 | 31 ± 9 | <0.001 |
| UL5-UL52-UL8 | pol α | (TCTG)15 | 5.5 ± 1.7 | NDc |
| UL5-UL52 | pol α | (TCTG)15 | 1.6 ± 0.9 | ND |
| UL5-UL52 + UL8 | pol α | (TCTG)15 | 3.3 ± 0.7 | ND |
a
Data are below the limit of detection.
b
In assays containing pol α-primase, the human primase dose generate small amounts of primers that pol α elongates, and primer transfer between human primase and pol α occurs intramolecularly (28). This small amount of coupled activity between the human primase and pol α would therefore confound attempts to identify direct primer transfer from herpes primase to pol α.
c
ND indicates no detectable products in the presence of binase.