Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Descriptive name | Genotype or description | Reference or source |
|---|---|---|---|
| Straina | |||
| SEY6210 | WT | MATα leu2-3,112 ura3-52 his3-Δ200 trp1-Δ901 lys2-801 suc2-Δ9 | Robinson et al. (1988) |
| BHY10 | WT CPY-I | SEY6210, CPY-INVERTASE::LEU2 ura3-52 his3-Δ200 trp1-Δ901 lys2-801 suc2-Δ9 | Horazdovsky et al. (1994) |
| MBY2 | vps4Δ CPY-I | BHY10, VPS4::TRP1 | Babst et al. (1997) |
| MBY3 | vps4Δ | SEY6210, VPS4::TRP1 | Babst et al. (1997) |
| MBY16 | vps4Δ vps36Δ | SEY62010.1, VPS4::TRP1, VPS36::HIS3 | Babst et al. (2002) |
| MBY28 | vps2Δ | SEY6210, VPS2::HIS3 | Babst et al. (2002) |
| BWY102 | vps24Δ | SEY6210, VPS24::HIS3 | Babst et al. (2002) |
| MBY37 | vps4Δ vps20Δ | MBY3, VPS20::HIS3 | Babst et al. (2002) |
| JPY50 | vps4Δ vta1Δ | MBY4, VTA1::HIS3 | Azmi et al. (2006) |
| MCY3 | vps4Δ ist1Δ | MBY3, IST1::HIS3 | Dimaano et al. (2008) |
| EEY1-3 | vps20Δ CPY-I | BHY10, VPS20::HIS3 | This study |
| EEY2-1 | vps20Δ | 6210, VPS20::HIS3 | Babst et al. (2002) |
| EEY8 | snf7Δ CPY-I | BHY10, SNF7::HIS3 | This study |
| EEY9 | snf7Δ | SEY6210, SNF7::HIS3 | Babst et al. (2002) |
| EEY12 | vps4Δ snf7Δ | MBY3, SNF7::HIS3 | Babst et al. (2002) |
| EEY26-1 | vps4Δ did2Δ | MBY3, DID2::HIS3 | Dimaano et al. (2008) |
| ASY4 | vps4Δ vps2Δ vps24 | 6210.1, VPS4::TRP1, VPS2::HIS3, VPS24::HIS3 | This study |
| ASY5 | vps2Δ vps24Δ | SEY6210, VPS24::HIS3, VPS2::HIS3 | This study |
| ASY8 | vps2Δ CPY-I | BHY10, VPS2::G418 | This study |
| ASY9 | vps24Δ CPY-I | BHY10, VPS24::G418 | This study |
| ASY12 | vps2Δ vps24Δ CPY-I | ASY8, VPS24::HIS3 | This study |
| ASY16 | vps4Δ vps20Δ snf7Δ | MBY37, SNF7::G418 | This study |
| ASY19 | snf7Δ vps20Δ CPY-I | EEY1-3, SNF7::G418 | This study |
| ASY20 | vps4Δ vps20Δ snf7Δ ist1Δ | ASY16, IST1::URA3 | This study |
| E. coli: XL1-blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10(tetr)] | Stratagene(La Jolla, CA) | |
| Plasmids | |||
| pAS22 | vps2(ΔC)-HA | URA3 (pRS416) vps2(ΔC)-HA | This study |
| pAS23 | vps24(ΔC)-HA | URA3 (pRS416) vps24(ΔC)-HA | This study |
| pAS28 | vps2(ΔC)-HA | LEU2 (pRS415) vps2(ΔC)-HA | This study |
| pAS42 | vps2(ΔC)-HA, vps24(ΔC)-HA | TRP1 (pRS414) vps2(ΔC)-HA, vps24(ΔC)-HA | This study |
| pAS44 | MIT-GFP | URA3 (pRS426) vps4(MIT)-GFP | This study |
| pAS47 | vps2(ΔC)-HA, vps24(ΔC)-HA | URA3 (pRS416) vps2(ΔC)-HA, vps24(ΔC)-HA | This study |
| pAS51 | GFP-vps4(ΔMIT(E233Q, S377A)) | URA3 (pRS416) GFP-vps4(ΔMIT(E233Q, S377A)) | This study |
| pAS58 | vps2(ΔC)-HA, vps24(ΔC)-HA | LEU2 (pRS415) vps2(ΔC)-HA, vps24(ΔC)-HA | This study |
| pAS59 | MIT(L64D)-GFP | URA3 (pRS426) vps4(MIT (L64D))-GFP | This study |
| pAS62 | vps4 (ΔMIT(E233Q)) | URA3 (pRS416) vps4(ΔMIT(E233Q)) | This study |
| pAS72 | snf7(L199D) | LEU2 (pRS415) snf(L199D) | This study |
| pAS74 | vps20(L188D) | LEU2 (pRS415) vps20(L188D) | This study |
| pAS76 | vps20(L188D)-HA | URA3 (pRS416) vps20(L188D)-HA | This study |
| pAS79 | vps4(S377A) | URA3 (pRS416) vps4(S377A) | This study |
| pMB4 | VPS4 | HIS3 (pRS413) VPS4 | Babst et al. (1997) |
| pMB66 | vps4(E233Q) | HIS3 (pRS413) vps4(E233Q) | Babst et al. (1998) |
| pMB168 | VPS20-HA | URA3 (pRS416) VPS20-HA | Babst et al. (2002) |
| pMB341 | GFP-vps4(ΔMIT(E233Q)) | URA3 (pRS416) P(CPY)-GFP-vps4(ΔMIT(E233Q)) | This study |
| pMB343 | vps4(E233Q)–GFP | URA3 (pRS416) vps4(E233Q)-HA-GFP | This study |
| pMB370 | vps4(L64D, E233Q)-GFP | URA3 (pRS416) vps4(L64D, E233Q)-HA-GFP | This study |
| pMB380 | vps4(I18D, E233Q)-GFP | URA3 (pRS416) vps4(I18D, E233Q)-HA-GFP | This study |
| pMB393 | snf7(L199D), vps20(L188D) | URA3 (pRS416) snf7(L199D), vps20(L188D) | This study |
| pMB394 | snf7(L199D), vps20(L188D) | LEU2 (pRS415) snf7(L199D), vps20(L188D) | This study |
| pPN3 | VPS20 | LEU2 (pRS415) VPS20 | This study |
| pVPS4(I18D) | vps4(I18D) | URA3 (pRS416) vps4(I18D) | This study |
| pVPS4(L64D) | vps4(L64D) | URA3 (pRS416) vps4(L64D) | This study |
| pVPS4(I18D, E233Q) | vps4(I18D, E233Q) | URA3 (pRS416) vps4(I18D, E233Q) | This study |
| pVPS4(L64D, E233Q) | vps4(L64D, E233Q) | URA3 (pRS416) vps4(L64D, E233Q) | This study |
| pGO45 | GFP-CPS | URA3(pRS426) GFP-CPS1 | Odorizzi et al. (1998) |
| pMC48 | MIT-GFP | URA3(pRS416) P(CPS1)-vps4(MIT)-GFP | This study |
| pMC50 | MIT(I18D)-GFP | URA3 (pRS416) P(CPS1)-vps4(MIT(I18D))-GFP | This study |
| pAH31 | GST-DID2 | (pGEX-KG) GST-DID2 | This study |
| pAH32 | GST-CT(DID2) | (pGEX-KG) GST-DID2(113-204) | This study |
| pMB411 | GST-vps20(C) | (pGEX-KG) GST-vps20(101-221) | This study |
| pMB412 | GST-snf7(C, L199D) | (pGEX-KG) GST-snf7(101-240)(L199D) | This study |
| pMB413 | GST-snf7(C) | (pGEX-KG) GST-snf7(101-240) | This study |
| pMB414 | GST-vps20(C, L188D) | (pGEX-KG) GST-snf7(101-221)(L188D) | This study |
| pAS85 | snf7(L199D), vps20(L188D), MIT-GFP | LEU2 (pRS415) snf7(L199D), vps20(L188D), P(CPS1)-MIT-GFP | This study |
a All strains are Saccharomyces cerevisiae are except the one marked E. coli.