Chicken embryo fibroblasts (CEF) when exposed to glucose-deficient culture medium developed 4- to 10-fold enhanced hexose transport activity within a few hours. Plasma membrane fractions prepared from starved and fed CEF revealed that starved cell membranes had a threefold greater glucose transport activity and [3H]cytochalasin B binding. The close correlation between transport activities of whole CEF and plasma membrane fractions indicates that hexose transport regulation during starvation results primarily in an increase in the number of functioning hexose transporters. The effect of protein synthesis inhibition on the overall process was studied with emetine, an inhibitor of translational elongation. Glucose-fed CEF treated with low concentrations of emetine (0.1 microM) showed a loss of transport greater than 65% within 4 h, but with higher concentrations of emetine (10 microM) there was no significant effect. Emetine treatment (0.1-10 microM) of CEF undergoing starvation virtually blocked any enhancement in transport whereas treatment of starved CEF led to only a slight loss of transport. Starved CEF refed with glucose had a decline of transport that was potentiated by low concentrations of emetine (0.1 microM); however, under these conditions high concentrations of emetine (10 microM) largely prevented loss of transport. Thus hexose transport regulation of CEF seems to reflect a balance between transporter synthesis and turnover. Transporter synthesis appears more sensitive to inhibition by emetine than turnover, whereas with hexose starvation there appears to be a decline in the activity of the transporter turnover process.