TABLE 2.
Plasmids used in this study
| Plasmid | Comment(s) | Reference |
|---|---|---|
| pUN100-LEU2-ProtA-TEV-NSP1-(591–823) (Nsil) | Construct contains an NsiI site changing amino acids 724 and 725 (VV) into (AL); called NSP1-C or Nsp1p-C | This study |
| pRS315-ADE2-GFP-NSP1-(591–823) | Like construct 1; ProtA replaced by GFP; called GFP-Nsp1p-C in Fig. 5 | This study |
| pUN100-LEU2-ProtA-nsp1 ts18 | See Materials and Methods; called nsp1 ts18 in Fig. 3, 4, and 5 | This study |
| pRS315-ADE2-GFP-nsp1 ts18 | Like construct 3; ProtA replaced by GFP; called GFP-nsp1 ts18 in Fig. 5 | This study |
| pUN100-LEU2-ProtA-TEV-nsp1-(591–775) | Called nsp1 tsΔ4 in Fig. 3 | This study |
| pSB32-LEU2-pADH-NSP1-L640S | Called nsp1-L640S in Fig. 3 and 4 | 44 |
| pUN100-LEU2-ProtA-TEV-NSP1-(630–823) (ΔN) | Called nsp1-(630–823) in Fig. 3 | This study |
| pUN100-LEU2-ProtA-TEV-NSP1-(638–823) | Called nsp1-(638–823) in Fig. 3 | This study |
| pUN100-LEU2-ProtA-TEV-NSP1-(642–823) | Called nsp1-(642–823) in Fig. 3 | This study |
| pSB32-LEU2-pADH-nsp1-ala6 | Called nsp1-ala6 in Fig. 3 and 4 | 3 |
| YCplac33-URA3-NUP159 (pLG4) | 8 | |
| pRS316-URA3-NUP82 | 19 | |
| pCH1122-URA3-ADE3-NIC96 | 20 | |
| pRS316-URA3-NUP57 | 20 | |
| pRS414-TRP1-GFP-NUP159-(2–1460) | Called GFP-Nup159p in Fig. 4 and 5 | 1 |
| pRS314-TRP1-GFP-NIC96-(2–838) | Called GFP-Nic96p in Fig. 5 | 1 |
| pRS314-TRP1-GFP-NUP57-(2–541) | Called GFP-Nup57p in Fig. 5 | This study |
| pRS314-TRP1-GFP-NUP82-(2–713) | Called GFP-Nup82p in Fig. 4 and 5 | 1 |
| pRS315-ADE2-GFP-NPL3 (283–397) | Called GFP-Npl3p in Fig. 4 | 39 |
| pGEX-4T-3 (Kan)-GST-NSP1-C (591–823) | Ampicillin resistance was released from pGEX-4T-3 and replaced by kanamycin resistance; called GST-Nsp1p-C in Fig. 1 and 2 | This study |
| pGEX-4T-3 (Amp)-GST-NSP1-(591–672) | Called GST-Nsp1p-coil 1 in Fig. 1 | This study |
| pGEX-4T-3 (Kan)-GST-NSP1-(673–781) | Called GST-Nsp1p-coil 2 + 3 in Fig. 1 | This study |
| pGEX-4T-3 (Kan)-GST-NSP1-(673–738) | Called GST-Nsp1p-coil 2 in Fig. 1 | This study |
| pGEX-4T-3 (Amp)-GST-NSP1-(591–738) | Called GST-Nsp1p-coil 1 + 2 in Fig. 1 | This study |
| pGEX-4T-3 (Amp)-GST-NSP1-(728–823) | Called GST-Nsp1p-coil 3 + 4 in Fig. 1 | This study |
| pET9d (Kan)-GST-NUP57-(2–541) | Called GST-Nup57p in Fig. 2 | This study |
| pET9d (Kan)-His6-NSP1-(591–823) | Called His6-Nsp1p-C in Fig. 2 | This study |
| pET9d (Kan)-His6-NUP159-(892–1460) | Called His6-Nup159p-C in Fig. 1 | This study |
| pET8c (Amp)-His6-NUP49-(2–472) | Called His6-Nup49p in Fig. 1 | 37 |
| pET8c (Amp)-His6-NUP57-(2–541) | Called His6-Nup57p in Fig. 1 and 2 | 37 |
| pET8c (Amp)-His6-NUP82-(2–713) | Called His6-Nup82p in Fig. 1 and 2 | 37 |