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. 2016 Mar 21;44(12):5629–5645. doi: 10.1093/nar/gkw168

Figure 6.

Figure 6.

Suppression of Snf1-T210 hyperphosphorylation requires functional Glc7. (A) Effect of Ssb1 and Bmh1 over-expression on the phosphorylation status of Snf1-T210 in a GLC7-T152K mutant strain. To induce the Glc7-T152K defect, cultures were shifted to 39°C for 1h prior to harvest. Aliquots of total cell extracts were analyzed via immunoblotting using the indicated antibodies. Sse1 served as a loading control. (B and C) Suppression of growth defects caused by the Glc7-T152K mutation. Serial 10-fold dilutions of the strains indicated were spotted onto YPD plates and were grown for 3 days at 39°C (B) or were spotted onto YPSuc plates containing 200 μg/ml 2-deoxy-glucose (2DG) and were incubated for 8 days at 30°C (C). (D) Snf1-T210 is not significantly hyperphosphorylated in Δsit4 cells. Aliquots of total cell extracts were analyzed via immunoblotting using the indicated antibodies. (E) Over-expression of Ssb or Bmh rescues lethality of a Δreg1Δsit4 strain. The strains indicated were grown on SD for 5 days, or on SD supplemented with 5-FOA for 6 days. For details compare ‘Materials and Methods’ section.