Table 1.
Quantified values for ZAP RNA binding mutant phenotypes.
| Isoform | Mutant | SINV RNA binding | ZAP-TRIM25 interaction | SINV replication inhibition | SINV translation inhibition | JEV translation inhibition |
|---|---|---|---|---|---|---|
| ZAPS | WT for ZnF mutants | 161.914 | 0.131 | 4.545 | 1.252 | 1.446 |
| ZAPS | C88R | 9.083 | 0.450 | 1.395 | 1.295 | 1.676 |
| ZAPS | H191R | 15.452 | 1.052 | 1.951 | N/A | 2.184 |
| ZAPS | WT for cavity mutants | 40.011 | 0.304 | 4.542 | 1.613 | 1.684 |
| ZAPS | V72A/Y108A/F144A | 1.444 | 0.792 | 1.777 | 1.033 | 1.598 |
| ZAPS | H176A/Y184A/R189A | 20.797 | 0.205 | 1.706 | 1.305 | 1.381 |
| ZAPS | C96A/Y98A | 0.408 | 0.550 | 1.361 | 1.166 | 1.260 |
| ZAPS | K107A/Y108A | 11.959 | 0.330 | 1.190 | 1.023 | 0.780 |
| ZAPS | E148A/K151A/R170A | 95.909 | 0.189 | 5.015 | N/A | 1.226 |
| ZAPL | WT for ZnF mutants | 93.414 | 0.055 | 12.652 | 1.574 | 2.416 |
| ZAPL | C88R | 0.034 | 1.126 | 2.756 | 1.253 | 5.191 |
| ZAPL | H191R | 0.761 | 0.899 | 3.043 | N/A | 9.046 |
| ZAPL | WT for cavity mutants | 281.812 | 0.076 | 18.090 | 1.580 | 2.444 |
| ZAPL | V72A/Y108A/F144A | 119.441 | 0.574 | 4.590 | 0.878 | 2.281 |
| ZAPL | H176A/F184A/R189A | 125.392 | 0.030 | 1.396 | 0.922 | 1.357 |
| ZAPL | C96A/Y98A | 68.060 | 0.329 | 4.774 | 1.257 | 3.551 |
| ZAPL | K107A/Y108A | 93.185 | 0.086 | 3.071 | 0.969 | 1.138 |
| ZAPL | E148A/K151A/R170A | 188.614 | 0.196 | 8.828 | N/A | 2.165 |
Immunoblots for ZAP SINV RNA binding assays and ZAP-TRIM25 co-IPs were quantified in ImageJ, with higher values indicating increased RNA or TRIM25 interaction. Fold inhibition values for viral inhibition assays were calculated relative to empty plasmid transfection, with higher values indicating greater viral replication or translation inhibition. N/A: SINV translation inhibition data on ZAP H191R and EKR mutants were not collected for analysis. Data are combined from two independent experiments.