TABLE 2.
RNA synthesis by recombinant RdRps using different GTP analogsa
| Analog | Mean % RNA synthesis ± SD
|
||||||||
|---|---|---|---|---|---|---|---|---|---|
| Recombinant RdRps
|
Replicases
|
||||||||
| HCV Δ21
|
BVDV Δ23
|
GBV Δ23
|
BMV | CMV (+Mn2+) | CCMV | ||||
| +Mn2+ | −Mn2+ | +Mn2+ | −Mn2+ | +Mn2+ | −Mn2+ | ||||
| Ribose | |||||||||
| GTP | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 |
| dGTP | 71 ± 8 | 0 ± 0 | 252 ± 3 | 1 ± 0 | 98 ± 1 | 0 ± 0 | 60 ± 2 | 73 ± 4 | 65 ± 15 |
| Phosphate | |||||||||
| GDP | 348 ± 29 | 117 ± 1 | 209 ± 40 | 4 ± 1 | 149 ± 2 | 50 ± 7 | 7 ± 0 | 69 ± 6 | 57 ± 7 |
| GMP | 535 ± 75 | 277 ± 30 | 234 ± 6 | 21 ± 2 | 154 ± 3 | 153 ± 8 | 4 ± 0 | 55 ± 5 | 50 ± 8 |
| Guanosine | 79 ± 12 | 0 ± 0 | 11 ± 3 | 0 ± 0 | 40 ± 6 | 2 ± 1 | NTb | NT | NT |
| GTP-γ-S | 85 ± 3 | 189 ± 16 | 116 ± 12 | 72 ± 2 | 43 ± 2 | 62 ± 5 | 120 ± 11 | 76 ± 6 | 36 ± 4 |
| GDP-β-S | 501 ± 50 | 173 ± 3 | 257 ± 23 | 9 ± 1 | 80 ± 9 | 27 ± 1 | 26 ± 0 | 78 ± 10 | 61 ± 4 |
| βγ-imido GTP | 118 ± 7 | 844 ± 90 | 394 ± 13 | 217 ± 26 | 136 ± 4 | 109 ± 4 | 50 ± 1 | 84 ± 8 | 69 ± 3 |
| Base | |||||||||
| XTP | 6 ± 3 | 2 ± 1 | 1 ± 1 | 0 ± 0 | 0 ± 0 | 0 ± 0 | 0 | 2 ± 3 | 1 ± 1 |
| ITP | 44 ± 8 | 1 ± 0.4 | 25 ± 1 | 1 ± 0 | 23 ± 1 | 1 ± 0 | 13 ± 2 | 73 ± 7 | 82 ± 8 |
| N7-meGTP | 87 ± 5 | 9 ± 4 | 61 ± 1 | 4 ± 0 | 128 ± 19 | 2 ± 0 | 12 ± 1 | 18 ± 3 | 19 ± 5 |
a
All reactions contained 4 mM Mg2+. Where indicated Mn2+ (+c), Mn2+ was added to a final concentration of 1 mM. LE19 is the template used for all of the RNA synthesis reactions using recombinant RdRps. The plant viral replicases used template −20/13, the subgenomic promoter for BMV. −Mn2+, no Mn2+ added.
b
NT, not tested.