Table 1.
Transcriptional activation by DNA-bound LexA-Snf1G53R
| Relevant genotype | Expressed protein | β-Galactosidase activity
|
|
|---|---|---|---|
| 4% Glu | 0.05% Glu | ||
| WT | LexA-Snf1 | <1 | <1 |
| WT | LexA-Snf1G53R | <1 | 240 |
| WT | LexA-Snf1G53R,K84R | <1 | <1 |
| snf4Δ | LexA-Snf1G53R | <1 | <1 |
| reg1Δ | LexA-Snf1G53R | 1,420 | 2,940 |
| reg1Δ | LexA | <1 | <1 |
| WT (pSH18-18) | LexA-Snf1G53R | 1 | 250 |
| mig1Δ (pSH18-18) | LexA-Snf1G53R | 2 | 440 |
Wild-type (WT) strain CTY10-5d (lexAop-lacZ) and snf4Δ3∷TRP1 or reg1Δ∷URA3 derivatives were transformed with pIT469, pRJ216, and pIT514, which express LexA fusions to Snf1, Snf1G53R, and Snf1G53R,K84R, respectively, and the vector pEG202 (34). Two strains of the S288C genetic background, wild-type FY250, and a mig1Δ mutant were transformed with both pRJ216 and the lexAop-lacZ reporter pSH18-18. Transformants were grown to mid-log phase in selective synthetic medium with 4% glucose (Glu) and were shifted to 0.05% glucose for 3 h. Values are average β-galactosidase activity for at least three transformants. Standard errors were <15%. Immunoblot analysis confirmed the stability of LexA-Snf1G53R in the snf4Δ strain.