TABLE 2.
Plasmids used in this study
| Plasmid | Description |
|---|---|
| pRS313 | CEN ARS HIS3 vector (26) |
| 313pBHB1–416 | Expresses a HA-BCY1 fusion, encoding HA-tagged wild-type Bcy1p, using the BCY1 promoter in vector pRS313; plasmid 313HBwt in Griffioen et al. (12) |
| 313pBHB125–416 | 313pBHB1–416 derivative encoding Bcy1 deleted for amino acids 1 to 124; plasmid 313HBΔN2 in Griffioen et al. (12) |
| 313pB(NLS)HBwt | 313pBHB1–416 derivative encoding Bcy1 fused with SV40 NLS |
| YCplac33 | CEN ARS URA3 vector (10) |
| 33AGHB1–124 | Expresses a HA-BCY1 fusion, encoding HA-tagged Bcy1 N terminus (amino acids 1 to 124) using the ADH1 promoter in vector YCplac33 |
| 33AGHB(1–124, Ser cluster I Ala) | 33AGHB1–124 derivative encoding Bcy1 bearing S3A, S4A, and S9A substitutions |
| 33AGHB(1–124, Ser cluster II Ala) | 33AGHB1–124 derivative encoding Bcy1 bearing S74A, S77A, S79A, S81A, S83A, and S84A substitutions |
| 33AGHB(1–124, Ser cluster I+II Ala) | 33AGHB1–124 derivative encoding Bcy1 bearing S3A, S4A, S9A, S74A, S77A, S79A, S81A, S83A, and S84A substitutions |
| 33pBGHBwt | Expresses a GFP-HA-BCY1 fusion, encoding wild-type Bcy1 fused with GFP-HA, using the BCY1 promoter in vector YCplac33 |
| 33pBGHB(Ser cluster I Ala) | 33pBGHBwt derivative encoding Bcy1 bearing S3A, S4A, and S9A substitutions |
| 33pBGHB(Ser cluster II Ala) | 33pBGHBwt derivative encoding Bcy1 bearing S74A, S77A, S79A, S81A, S83A, and S84A substitutions |
| 33pBGHB(Ser cluster I+II Ala) | 33pBGHBwt derivative encoding Bcy1 bearing S3A, S4A, S9A, S74A, S77A, S79A, S81A, S83A, and S84A substitutions |
| 195A2 | 2μm, ADE2 vector (this study) |
| 195A2-pBGHBwt | Expresses a GFP-HA-BCY1 fusion, encoding wild-type Bcy1p fused with GFP-HA, using the BCY1 promoter in vector 195ADE2 |
| 195A2-pBGHB(Ser cluster I Asp) | 195A2-pBGHBwt derivative encoding Bcy1 bearing S3D, S4D, and S9D substitutions |
| 195A2-BGHB(Ser cluster II Asp) | 195A2-pBGHBwt derivative encoding Bcy1 bearing S74D, S77D, S79D, S81D, S83D, and S84D substitutions |
| 195A2-pBGHB(Ser cluster I+II Asp) | 195A2-pBGHBwt derivative encoding Bcy1 bearing S3D, S4D, S9D, S74D, S77D, S79D, S81D, S83D, and S84D substitutions |
| pRS316 | CEN URA3 vector (10) |
| 316pADH1-ZDS1 | Expresses ZDS1, using the ADH1 promoter, in vector pRS316 |
| pGBT9 | Two-hybrid expression vector with GAL4 DNA-binding domain (DBD) (15) |
| T9-BCY1wt | BCY1 inserted in pGBT9; expresses Ga14-DBD fused Bcy1 |
| T9-BCY1125–416 | T9-BCY1wt derivative encoding Bcy1 deleted for amino acids 1 to 124 |
| T9-BCY11–179 | T9-BCY1wt derivative encoding Bcy1 deleted for amino acids 180 to 416 |