CYTOCHALISIN-D EXERTS STIMULATORY AND INHIBITORY EFFECTS ON INSULIN-INDUCED GLUCOKINASE MESSENGER-RNA EXPRESSION IN HEPATOCYTES

The microfilament cytoskeleton is postulated to have a role in the localization, transport and anchorage of certain specific mRNAs. We investigated the effects of cytochalasin D, a fungal metabolite that binds to actin and disrupts the microfilament structure, on insulin-induced expression of glucokinase mRNA in rat hepatocyte cultures. Cytochalasin-D significantly potentiates insulin-induced glucokinase mRNA expression at 100 nM concentration but counteracts glucokinase expression at 2-20 mu M. The latter effect is at least in part due to an increase in glucokinase mRNA degradation. This effect of cytochalasin D cannot be accounted for by an increase in cAMP and is also not due to a non-specific effect on mRNA degradation since albumin mRNA levels were not affected by cytochalasin-D and actin mRNA and tubulin mRNA levels were increased. Measurement of glucokinase mRNA release from digitonin-permeabilized hepatocytes suggests that cytochalasin D does not cause acute dissociation of glucokinase mRNA from its binding site. The increased degradation of glucokinase mRNA suggests involvement of the cytoskeleton in glucokinase mRNA stability. However, an additional effect of cytochalasin D on the insulin signalling mechanism cannot be excluded.