TABLE 1.
Summary of the effects of the mutations tested for RNA accumulation in barley protoplasts
| Template | Construct (sequence) | % of wild-type levela
|
||
|---|---|---|---|---|
| Minus-strand RNA3 | Plus-strand RNA3 | Plus-strand RNA4 | ||
| Minus-strand RNA3 | Wild type (. . .uaaaagagacca3′) | 100 | 100 | 100 |
| M1,2ΔCA (. . .uaaaagagac--3′) | 59 ± 5 | 33 ± 1 | 36 ± 1 | |
| M1,2CC/GG (. . .uaaaagagaGGa3′) | 99 ± 4 | 83 ± 11 | 69 ± 9 | |
| M1C/G (. . .uaaaagagacGa3′) | 114 ± 1 | 108 ± 4 | 121 ± 4 | |
| M2C/G (. . .uaaaagagaGca3′) | 90 ± 14 | 109 ± 3 | 169 ± 7 | |
| M2C/A (. . .uaaaagagaAca3′) | 104 | 80 | 105 | |
| M2C/U (. . .uaaaagagaUca3′) | 85 | 53 | 71 | |
| M3A/C (. . .uaaaagagCcca3′) | 97 | NT | NT | |
| M3A/G (. . .uaaaagagGcca3′) | 56 | 43 | 51 | |
| M3A/U (. . .uaaaagagUcca3′) | 113 | 91 | 98 | |
| M5A/G (. . .uaaaagGgacca3′) | 86 | 63 | 74 | |
| M7A/G (. . .uaaaGgagacca3′) | 128 | 99 | 105 | |
| Plus-strand RNA3 | Wild type (3′cauuuuaugguu. . .) | 100 | 100 | 100 |
| P2A/C (3′ cCuuuuaugguu. . .) | 74 ± 5 | 10 ± 3 | 216 ± 14 | |
| P2A/U (3′ cUuuuuaugguu. . .) | 68 ± 2 | 9 ± 1 | 109 ± 18 | |
| P2A/G (3′ cGuuuuaugguu. . .) | 40 ± 4 | 6 ± 1 | 18 ± 2 | |
| P3U/C (3′ caCuuuaugguu. . .) | 52 ± 4 | 23 ± 4 | 10 ± 5 | |
| P3U/G (3′ caGuuuaugguu. . .) | 89 ± 8 | 18 ± 7 | 345 ± 28 | |
| P3U/A (3′ caAuuuaugguu. . .) | 90 ± 2 | 24 ± 4 | 358 ± 34 | |
| P4U/G (3′ cauGuuaugguu. . .) | 68 ± 4 | 62 ± 3 | 103 ± 2 | |
| P4U/A (3′ cauAuuaugguu. . .) | 66 ± 4 | 50 ± 3 | 154 ± 2 | |
| P4U/C (3′ cauCuuaugguu. . .) | 44 ± 3 | 35 ± 2 | 136 ± 2 | |
| P5U/G (3′ cauuGuaugguu. . .) | 106 ± 1 | 58 ± 4 | 99 ± 11 | |
| P5U/C (3′ cauuCuaugguu. . .) | 85 ± 1 | 50 ± 5 | 156 ± 6 | |
| P5U/A (3′ cauuAuaugguu. . .) | 72 ± 11 | 64 ± 3 | 128 ± 24 | |
| P7A/C (3′ cauuuuCugguu. . .) | 65 ± 5 | 50 ± 7 | 47 ± 11 | |
| P7A/U (3′ cauuuuUugguu. . .) | 61 ± 1 | 50 ± 7 | 55 ± 7 | |
| P11U/C (3′ cauuuuauggCu. . .) | 32 ± 7 | 32 ± 4 | 28 ± 2 | |
| P11U/A (3′ cauuuuauggAu. . .) | 48 ± 1 | 60 ± 4 | 46 ± 7 | |
| Subgenomic RNA | Wild type (3′. . .cauaauuau. . .) | 100 | 100 | 100 |
| S1C/U (3′. . .Uauaauuau. . .) | 48 ± 3 | 25 ± 3 | 1 ± 1 | |
| S2A/C (3′. . .cCuaauuau. . .) | 52 ± 2 | 28 ± 2 | 1 ± 1 | |
| S2A/G (3′. . .cGuaauuau. . .) | 69 ± 5 | 48 ± 8 | 8 ± 1 | |
| S2A/U (3′. . .cUuaauuau. . .) | 62 ± 3 | 56 ± 1 | 9 ± 1 | |
| S3U/C (3′. . .caCaauuau. . .) | 42 ± 2 | 42 ± 5 | 14 ± 1 | |
| S3U/G (3′. . .caGaauuau. . .) | 35 ± 1 | 36 ± 4 | 7 ± 1 | |
| S3U/A (3′. . .caAaauuau. . .) | 52 ± 3 | 72 ± 2 | 13 ± 1 | |
| S5A/C (3′. . .cauaCuuau. . .) | 37b | 33b | 16b | |
| S5A/G (3′. . .cauaGuuau. . .) | 46 ± 1 | 60 ± 1 | 48 ± 1 | |
| S5A/U (3′. . .cauaUuuau. . .) | 114 ± 9 | 155 ± 1 | 85 ± 8 | |
| S7U/C (3′. . .cauaauCau. . .) | 59 ± 1 | 59 ± 3 | 25 ± 3 | |
| S7U/A (3′. . .cauaauAau. . .) | 72 ± 1 | 81 ± 4 | 72 ± 6 | |
| S8A/C (3′. . .cauaauuCu. . .) | 31 ± 1 | 36 ± 2 | 25 ± 2 | |
| S8A/G (3′. . .cauaauuGu. . .) | 41 ± 2 | 34 ± 1 | 12 ± 1 | |
| S8A/U (3′. . .cauaauuUu. . .) | 70 ± 11 | 73 ± 10 | 66 ± 5 | |
| SIWT (3′. . .cauaauuaAAUUAu. . .) | 100 | 100 | 100 | |
| SI10′A/C (3′. . .cauaauuaACUUAu. . .) | 100 | 115 | 124 | |
| SI10′A/G (3′. . .cauaauuaAGUUAu. . .) | 75b | 86b | 63b | |
| SI10′A/U (3′. . .cauaauuaAUUUAu. . .) | 110b | 171b | 165b | |
| SI12′U/A (3′. . .cauaauuaAAUAAu. . .) | 101 | 86 | 97 | |
| SI12′U/C (3′. . .cauaauuaAAUCAu. . .) | 110 | 114 | 185 | |
| SI13′A/C (3′. . .cauaauuaAAUUCu. . .) | 143 | 170 | 253 | |
| SI13′A/G (3′. . .cauaauuaAAUUGu. . .) | 162 | 182 | 252 | |
| SI13′A/U (3′. . .cauaauuaAAUUUu. . .) | 134 | 171 | 199 | |
a
Numbers in bold denote that the mutation is in the template for a particular mode of synthesis. Where standard deviations are not shown, the data were from only two independently assayed samples. NT, not tested.
b
These values were adjusted to normalize for the amount of rRNA in these samples.