Table 1.
Strains and plasmids
| Strains and plasmids | Description | Source |
|---|---|---|
| Yeast strains | ||
| L40 | MATa trp1 leu2 his3 LYS2::(lexAop)4-HIS3 URA3::(lexAop)4-lacZ | Vojtek et al., 1993 |
| SEY6210 | MATα leu2-3,112 ura3-52 his3-Δ200 trp1Δ901 lys2-801 suc2-Δ9 | Robinson et al., 1988 |
| BHY10 | SEY6210; leu2-3, 112::pBHY11(CPY-Inv LEU2) | Horazdovsky et al., 1994 |
| CCY120 | SEY6210; vps45Δ2::HIS3 | Cowles et al., 1994 |
| PHY102 | SEY6210; vps34Δ1::TRP1 | Herman and Emr, 1990 |
| CBY31 | SEY6210; Δpep12::HIS3 | Burd et al., 1997 |
| DDY3477 | SEY6210; vps34ts | This study |
| GTY102 | SEY6210; vac1Δ2::URA3 | This study |
| GTY104 | SEY6210; vac1Δ3::NEO | This study |
| GTY105 | BHY10; vac1Δ3::NEO | This study |
| GTY106 | SEY6210; vsp21::HIS3 vac1Δ3::NEO | This study |
| GTY107 | SEY6210; vps45Δ2::HIS3 vac1Δ3::NEO | This study |
| GTY108 | SEY6210; Δpep12::HIS3 vac1Δ3::NEO | This study |
| GTY109 | SEY6210; vps34Δ1::TRP1 vac1Δ3::NEO | This study |
| GTY110 | SEY6210; vsp34ts vac1Δ3::NEO | This study |
| GTY111 | SEY6210; Δpep12::HIS3 vac1Δ3::NEO vps34Δ1::TRP1 | This study |
| GTY112 | BHY10; vac1Δ2::URA3 vps21Δ3::NEO | This study |
| E. coli strains | ||
| XL1Blue | (recA1 endA1 gyrA96 thi-1 hsdR17 supE44) | Stratagene |
| M15 | (NalS StrS rifS lac− ara− gal− mtl− F− recA+ uvr+) | Qiagen |
| Plasmids | ||
| pVJL11 | Bait Vector | Jullien-Flores et al., 1995 |
| pGADGH | Prey Vector | Hannon et al., 1993 |
| pSM491 | Triple HA-tag with NotI linkers in pBS | Obtained from Pamela A. Marshall and Joel M. Goodman |
| pRS series Vectors | yeast expression vectors | Christianson et al., 1992 |
| pBS | pBluescript II | Stratagene |
| pQE31 | Histidine-tagged E. coli expression vector | Qiagen |
| pGT21-1 | Wild-type Vps21 bait in pVJL11 | Hama et al., 1999 |
| pGT21-2 | S21N-Vps21p bait in pVJL11 | Hama et al., 1999 |
| pGT21-3 | Q66L-Vps21p bait in pVJL11 | This study |
| pGT21-4 | Q66L-vps21 in pQE31 | This study |
| pBHY21-10 | VPS21 in pRS414 | Horazdovsky et al., 1994 |
| pBHY21-13 | Q66L-VPS21 in pRS414 | This study |
| pBHY21-23 | Q66L-VPS21 in pRS424 | This study |
| pBHY21-49 | VPS21-49 allele in pRS416 | This study |
| pBHY21-79 | Wild-type VPS21 in pQE31 | This study |
| pLW102 | vac1Δ2::URA3 in pRS316 | Weisman and Wickner, 1992 |
| pGT102 | vac1Δ2::URA3 in pBS | This study |
| pGT104 | vac1Δ3::NEO in pBS | This study |
| pGT1-1 | Wild-type VAC1 in pRS415 | This study |
| pVAC1-425 | Wild-type VAC1 in pRS425 | Weisman and Wickner, 1992 |
| pGT1-2 | Wild-type VAC1 in pRS413 | This study |
| pGT1-3 | Wild-type VAC1 in pRS423 | This study |
| pGT1-4 | C221S-vac1 in pRS413 | This study |
| pGT1-5 | C237S-vac1 in pRS415 | This study |
| pGT1-6 | C292S-vac1 in pRS413 | This study |
| pGT1-7 | C221/292S-vac1 in pRS413 | This study |
| pGT1-8 | Wild-type Vac1 bait in pVJL11 | This study |
| pGT1-9 | Wild-type Vac1 prey in pGADGH | This study |
| pGT1-10 | C221S-Vac1p prey in pGADGH | This study |
| pGT1-11 | C292S-Vac1p prey in pGADGH | This study |
| pGT1-12 | C221/292S-Vac1p prey in pGADGH | This study |
| pGT1-16 | VAC1 w/C-terminal HA-tag in pRS413 | This study |
| pGT1-17 | VAC1 w/C-terminal HA-tag in pRS415 | This study |
| pGT1-18 | C221S-VAC1 w/C-terminal HA-tag in pRS415 | This study |
| pGT1-19 | C292S-VAC1 w/C-terminal HA-tag in pRS415 | This study |
| pGT1-20 | C221/292S-VAC1 w/C-terminal HA-tag in pRS415 | This study |
| pGT1-21 | VAC1 w/C-terminal HA-tag in pRS423 | This study |
| pGT45-1 | Vps45p bait in pVJL11 | This study |
| pGT45-2 | Vps45p prey in pGADGH | This study |
| pVPS45-1 | original VPS45 genomic clone | Cowles et al., 1994 |
| pBHY45-CC2 | VPS45 in pRS414 | This study |
| pBHY45-3 | VPS45 in pRS416 | This study |
| pBHY45-4 | VPS45 in pRS426 | This study |
| pBHY45-7 | vps45 ts mutant in pRS414 | This study |