FIGURE 1.

Snf1 Thr-210 phosphorylation in mutants lacking Ptc1 and other PP2C phosphatases. Cells of the indicated relevant genotype were collected after growth on 2% (H, high) glucose or after a shift to 0.05% (L, low) glucose for 10 min. For B, aliquots of cultures shifted to low glucose were replenished with 2% glucose for 2.5 or 5 min, as indicated. Protein extracts were separated by SDS-PAGE and subjected to immunoblot analysis with anti-Thr(P)-172-AMPK to detect phosphorylated Thr-210 (pT210) and anti-Myc (A and B) or anti-polyhistidine (C) to detect Snf1 proteins. A and B, independent transformants expressed Snf1-myc from the SNF1 promoter on a centromeric plasmid. B, analysis of snf1Δ and ptc1Δ snf1Δ cells (with wild-type GLC3) gave results similar to those shown. C, cells carrying mutations ptc1Δ through ptc7Δ expressed Snf1 from the genomic locus and Snf1(1–309)-myc from pYL411. All lanes are from the same blot. Arrows mark the positions of Snf1 and Snf1(1–309).