TABLE 1.
YFP/RFP ratios of constructs in S. cerevisiae, S. pombe, and mammalian cells (HEK293)
The median of YFP/RFP ratio for each construct was calculated from 10,000 cells collected in one flow cytometry run and used to indicate the abundance of the YFP substrate in yeast cells as described under “Experimental Procedures.” Data represent mean values and standard errors determined from at least three repeat experiments. ND is not determined; NS indicates derived from influenza A virus non-structural protein 1 (NS1); GRR indicates glycine-rich region (derived from human p105); NB indicates derived from influenza B virus glycoprotein NB; SNS indicates tandem repeat of SP2-NB-SP2; NBS indicates tandem repeat of NB-NB-SP2; DRR indicates aspartic acid (D)-rich region (derived from S. cerevisiae Cdc34); SP1 indicates peptide region 1 in influenza A virus M2 protein used to produce antisera; SP2 indicates peptide region 2 in influenza A virus M2 protein used to produce antisera; SPmix indicates tandem repeat of SP1 and SP2 (SP2-SP1-SP2-SP1-SP2); PEST indicates sequence from human IκBα; SRR indicates serine-rich region (derived from herpes virus 1 ICP4); Su9, derived from subunit 9 of N. crassa ATP synthase component Fo; eRR, derived from E. coli lacI; ODC indicates derived from ornithine decarboxylase; 35 indicates derived from S. cerevisiae cytochrome b2.
| Tail | Name |
S. cerevisiae |
S. pombe | HEK293 | ||
|---|---|---|---|---|---|---|
| pACT1 UbL-YFP-tail pACT1 dsRed | pTPI1 UbL-YFP-tail pPGK1 dsRed | pTPI1 UbL-GFP-tail pPGK1 dsRed | ||||
| A | 35 | 0.15 ± 0.01 | 0.12 ± 0.02 | 0.13 ± 0.01 | 0.73 ± 0.06 | 0.20 ± 0.02 |
| B | ODC | 0.13 ± 0.01 | 0.13 ± 0.01 | 0.034 ± 0.004 | 1.0 ± 0.3 | 0.13 ± 0.02 |
| C | Poly(G) | 1.00 ± 0.05 | 1.22 ± 0.05 | 1.07 ± 0.08 | ND | 0.51 ± 0.01 |
| D | GRR | 1.2 ± 0.1 | 3.9 ± 0.1 | 1.9 ± 0.1 | 4.8 ± 0.2 | 0.83 ± 0.05 |
| E | SRR | 2.7 ± 0.2 | 4.6 ± 0.1 | 3.81 ± 0.09 | 6.54 ± 0.07 | 0.87 ± 0.02 |
| F | NB | 0.27 ± 0.01 | 0.79 ± 0.07 | 0.27 ± 0.02 | 2.8 ± 0.2 | 0.31 ± 0.02 |
| G | NS | 0.54 ± 0.03 | 1.12 ± 0.09 | 0.80 ± 0.03 | 2.3 ± 0.1 | 0.27 ± 0.02 |
| H | SP1 | 0.48 ± 0.01 | 0.88 ± 0.08 | 0.63 ± 0.01 | 3.0 ± 0.1 | 0.47 ± 0.03 |
| I | SP2 | 2.2 ± 0.2 | 6.0 ± 0.1 | 4.30 ± 0.06 | 5.65 ± 0.07 | 0.86 ± 0.04 |
| J | SPmix | 1.7 ± 0.1 | 3.5 ± 0.3 | 2.40 ± 0.07 | 5.94 ± 0.09 | 0.834 ± 0.004 |
| K | SNS | 1.29 ± 0.07 | 4.4 ± 0.2 | 2.7 ± 0.2 | 4.77 ± 0.07 | 0.70 ± 0.02 |
| L | NBS | 0.62 ± 0.02 | 2.3 ± 0.2 | 1.01 ± 0.07 | 5.2 ± 0.1 | 0.54 ± 0.02 |
| M | DRR | 2.2 ± 0.2 | 5.4 ± 0.1 | 4.6 ± 0.3 | 4.9 ± 0.2 | 0.76 ± 0.03 |
| N | eRR | 0.43 ± 0.02 | 0.70 ± 0.02 | 0.40 ± 0.01 | 1.10 ± 0.05 | 0.28 ± 0.02 |
| O | PEST | 0.71 ± 0.01 | 1.47 ± 0.04 | 0.98 ± 0.08 | 2.85 ± 0.08 | 0.21 ± 0.01 |
| P | Su9 | 0.11 ± 0.03 | 0.19 ± 0.01 | 0.14 ± 0.02 | 1.7 ± 0.2 | 0.16 ± 0.01 |
| Q | No tail | 2.5 ± 0.2 | 8.3 ± 0.2 | 5.1 ± 0.3 | 6.8 ± 0.1 | 0.83 ± 0.06 |