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J. Biol. Chem., Vol. 265, Issue 3, 1516-1523, Jan, 1990

Glucose transport activity in L6 muscle cells is regulated by the coordinate control of subcellular glucose transporter distribution, biosynthesis, and mRNA transcription

PS Walker, T Ramlal, V Sarabia, UM Koivisto, PJ Bilan, JE Pessin and A Klip
Department of Physiology and Biophysics, University of Iowa, Iowa City 52242.

Chronic (24 h) insulin treatment and/or glucose deprivation of differentiated rat L6 skeletal muscle cells resulted in an increase in glucose transport activity and a 2-3-fold increase in the number of plasma membrane-associated cytochalasin B binding sites and immunoreactive glucose transporters. In contrast to the acute effect of insulin, chronic treatment did not decrease the number of cytochalasin B binding sites or immunoreactive glucose transporter proteins present in intracellular low density microsomes. Although acute insulin stimulation of glucose transport activity was not affected by cycloheximide, chronic insulin stimulation of glucose transport activity and glucose transporter protein were decreased. In contrast, the stimulation of glucose transport activity by both acute and chronic glucose deprivation were cycloheximide-insensitive. Previously we have reported that chronic insulin treatment transiently induces the rat brain/HepG2 glucose transporter subtype (GLUT-1) mRNA, whereas glucose deprivation induces a substained increase (Walker, P. S., Ramlal, T., Donovan, J. A., Doering, T. P., Sandra, A., Klip, A., and Pessin, J. E. (1989) J. Biol. Chem. 264, 6587-6595). Consistent with these data, nuclear run-on analysis demonstrated a transient 3-fold increase in the rate of GLUT-1 glucose transporter mRNA transcription induced by either chronic insulin treatment or glucose deprivation. The combination of chronic insulin treatment with glucose deprivation resulted in a more persistent 3-4-fold increase in transcription rate than either treatment alone. These data demonstrate that prolonged insulin- and glucose-dependent regulation of glucose transporter function occurs by a complex mechanism which includes enhanced GLUT-1 mRNA transcription and glucose transporter synthesis, as well as changes in the subcellular distribution of glucose transporter proteins.
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