TABLE 1.
Summary of interactions between the viroplasmic proteins NSP2 and NSP5 and host proteinsa
| Host protein | Interaction by mass spectrometry |
Interaction by PD-Western blotting |
Colocalization with VM | Total protein level (8 hpi) | ||
|---|---|---|---|---|---|---|
| NSP5 | NSP2 | NSP5 | NSP2 | |||
| hnRNPs | ||||||
| hnRNP A1 | + | − | + | +c | − | ↑ |
| hnRNP C1/2 | + | − | + | + | + | ↑ |
| hnRNP D | + | + | + | + | + | ↓ |
| hnRNP E | + | + | + | + | + | ↑ |
| hnRNP F | + | + | + | + | + | ↑ |
| hnRNP H | + | + | + | + | + | ↑ |
| hnRNP I | + | + | + | + | + | ↑ |
| hnRNP K | + | + | + | + | + | ↑ |
| hnRNP L | + | + | + | + | + | ↑ |
| hnRNP M | + | − | + | + | + | ± |
| hnRNP Q | − | − | − | − | − | ↑ |
| hnRNP U | + | + | + | + | + | ↑ |
| ARE-BPs | ||||||
| BRF1 | − | − | + | +b | + | ↓ |
| HuR | − | − | − | +c | + | ↑ |
| hnRNP D | + | + | + | + | + | ↓ |
| KSRP | − | − | NT | NT | + | ↑ |
| Staufen | − | − | + | + | + | ± |
| TIA1 | − | − | + | + | + | ↑ |
| TIAL-1 | − | − | − | +b | + | ↓ |
| TTP | − | − | NT | NT | + | ± |
| Other nuclear/cytoplasmic proteins | ||||||
| RPS8 | + | − | + | − | + | ↑ |
| VPS35 | + | + | + | − | + | ↑ |
| Sec31A | + | + | + | + | + | ↑ |
| α/β-Tubulin | − | + | NT | NT | NT | ± |
| G3BP1 | − | − | − | − | − | ↓ |
| Transport proteins | ||||||
| Transportin1 | − | − | − | − | + | ↓ |
| Exportin1 | − | − | + | − | + | ↑ |
| Importin-β | − | − | − | + | + | ↑ |
| Ran | − | − | − | + | + | NT |
The interactions identified between the cellular proteins and viral proteins NSP2 and NSP5 by a PD assay followed by immunoblotting and colocalization by ICM (Fig. 1a and b and 4) are summarized. +, positive interaction/colocalization; −, no interaction/colocalization. ↑ and ↓ indicate increased and decreased host protein levels in virus-infected cells in comparison to those in uninfected cells, respectively; ± indicates that there was no significant change in the protein levels between virus-infected and uninfected cells. NT, not tested.
Loss of binding of NSP2 with BRF1 and TIAL-1 in RNase-treated cell extracts.
Reduced binding of NSP2 with hnRNP A1 and HuR in RNase-treated cell extracts.