Fig. 1.

Glucose regulation of Thr-210 phosphorylation in the absence of the heterotrimer. Cells of the indicated genotype were grown on high (H) glucose, resuspended in low (L) glucose for 10 min, and replenished with 2% glucose (+G) for 15 min. Protein extracts were separated by SDS/PAGE and subjected to immunoblot analysis to detect phosphorylated Thr-210 (pT210). Membranes were reprobed to detect Snf1 polypeptides. (A) Differing amounts of protein extract were loaded, and Snf1 was detected with anti-polyhistidine. (B–D) Cells expressed Snf1-myc or Snf1(1–309)-myc from the native promoter on centromeric plasmids. Protein extract (4 μg) was used. (B) Lanes are from the same blot but longer exposures are shown for Snf1-myc. (D) SNF1-myc cells expressed Snf1-8xmyc from the genomic locus.