Fig. 4

Activation of Ras by Fru1,6bisP. a Ras-GTP level 3 min after addition of 5 mM of different glycolytic metabolites to permeabilized yeast spheroplasts. Glu6P, glucose-6-phosphate; Tre6P, trehalose-6-phosphate; Fru6P, fructose-6-phosphate; Fru1,6bisP, fructose-1,6-bisphosphate; DHAP, dihydroxyacetone phosphate; GAP, glyceraldehyde-3-phosphate; 3PG, 3-phosphoglycerate; PEP, phosphoenolpyruvate. Quantification of the signals as Ras-GTP level/total Ras level compared to the ratio for the water control set at 1.0. The values were determined through quantification by densitometry and are expressed as relative arbitrary units. b Activation of Ras in permeabilized spheroplasts with different concentrations of GTP, Fru1,6bisP, DHAP and GAP. Quantification of the signals as Ras-GTP level/total Ras level compared to the ratio for the water control set at 1.0. The values were determined through quantification by densitometry and are expressed as relative arbitrary units. c Ras-GTP level before and 10 min after addition of 100 mM glucose (glu) to cells grown on glycerol +2.5 mM glucose (gly). Quantification of the signals as Ras-GTP level/total Ras level compared to the ratio for tps1∆ on glycerol set at 1.0. The values were determined through quantification by densitometry and are expressed as relative arbitrary units. d Intracellular ATP, Glu6P, and Fru1,6bisP concentration before and after addition of 100 mM glucose to cells grown in glycerol +2.5 mM glucose