TABLE 1.
EAV nsp10 mutants and their phenotypes
| Construct | Substitution | Codon
|
Result ofa:
|
Description of phenotypeb | |||
|---|---|---|---|---|---|---|---|
| Wild type | Mutant | nsp3 IFA | N protein IFA | Infectivity | |||
| EAV030H | None (wild-type) | + | + | + | Wild type | ||
| C2374H | Cys-2374→His | UGC | CAC | − | − | − | Nonviable |
| C2377H | Cys-2377→His | UGU | CAU | − | − | − | Nonviable |
| K2384A | Lys-2384→Ala | AAG | GCG | − | − | − | Nonviable |
| S2385A | Ser-2385→Ala | UCU | GCU | + | + | + | Wild type |
| S2385T | Ser-2385→Thr | UCU | ACU | + | + | + | Wild type |
| C2387H | Cys-2387→His | UGU | CAU | − | − | − | Nonviable |
| G2388A | Gly-2388→Ala | GGA | GCA | + | + | + | Viable, delayed |
| G2388V | Gly-2388→Val | GGA | GUA | + | + | + | Viable, delayed |
| C2392H | Cys-2392→His | UGU | CAU | − | − | − | Nonviable |
| C2395H | Cys-2395→His | UGU | CAC | + | + | + | Viable, delayed |
| H2399C | His-2399→Cys | CAC | UGC | − | − | − | Nonviable |
| H2402C | His-2402→Cys | CAU | UGU | − | − | − | Nonviable |
| C2403H | Cys-2403→His | UGU | CAU | − | − | − | Nonviable |
| H2404C | His-2404→Cys | CAC | UGU | − | − | − | Nonviable |
| CCH→HHC | Cys-2395→His | UGU | CAC | − | − | − | Nonviable |
| Cys-2403→His | UGU | CAU | |||||
| His-2404→Cys | CAC | UGC | |||||
| C2412H | Cys-2412→His | UGC | CAU | − | − | − | Nonviable |
| H2414C | His-2414→Cys | CAC | UGC | + | + | − | RNA synthesis, but not viable |
| H2414Y | His-2414→Tyr | CAC | UAC | − | − | − | Nonviable |
| D2415A | Asp-2415→Ala | GAC | GCC | − | − | − | Nonviable |
| R2419A | Arg-2419→Ala | CGC | GCG | + | + | + | Viable, delayed |
| S2420A | Ser-2420→Ala | UCC | GCU | + | + | + | Wild type |
| S2420T | Ser-2420→Thr | UCC | ACU | + | + | + | Viable, cold sensitive |
| C2423H | Cys-2423→His | UGC | CAC | − | − | − | Nonviable |
| C2426H | Cys-2426→His | UGU | CAU | − | − | − | Nonviable |
| C2426N | Cys-2426→Asn | UGU | AAU | − | − | − | Nonviable |
| E2427A | Glu-2427→Ala | GAG | GCG | + | + | + | Viable, delayed |
| ΔE2427 | Glu-2427 deletion | GAG | − | − | − | Nonviable | |
| ΔG2428 | Gly-2428 deletion | GGU | − | − | − | Nonviable | |
| EAV030F | Ser-2429→Pro | UCC | CCC | + | − | − | Defect in sg mRNA synthesis |
| S2429A | Ser-2429→Ala | UCC | GCC | + | + | + | Wild type |
| S2429C | Ser-2429→Cys | UCC | UGC | + | + | + | Wild type |
| S2429G | Ser-2429→Gly | UCC | GGC | + | + | + | Wild type |
| S2429H | Ser-2429→His | UCC | CAC | + | + | + | Wild type |
| S2429L | Ser-2429→Leu | UCC | CUC | + | + | + | Wild type |
| S2429T | Ser-2429→Thr | UCC | ACC | + | + | + | Wild type |
| S2429G/P2430G | Ser-2429→Gly | UCC | GGC | + | ± | − | Defect in sg mRNA synthesis |
| Pro-2430→Gly | CCA | GGA | |||||
| S2429P/P2430S | Ser-2429→Pro | UCC | CCC | + | − | − | Defect in sg mRNA synthesis |
| Pro-2430→Ser | CCA | UCA | Delayed genome replication | ||||
a
+, positive; −, negative; ±, intermediate.
b
Delayed, replication delayed compared to that of wt virus at the same temperature, as judged by IFA (onset of nonstructural and structural protein synthesis) and virus spread to untransfected cells