Evidence of two mechanisms for the activation of the glucose transporter GLUT1 by anisomycin: p38(MAP kinase) activation and protein synthesis inhibition in mammalian cells.
1. Inhibitors of protein synthesis stimulate sugar transport in mammalian cells through activation of plasma membrane GLUT1, the housekeeping isoform of the glucose transporter. However, it has been reported that some of these compounds, in addition to their effect on protein synthesis, also activate protein kinases. 2. In the present study we have explored the role of these two effects on GLUT1 activation. In 3T3-L1 adipocytes and Clone 9 cells, stimulation of sugar transport by puromycin, a translational inhibitor that does not activate kinases, was not detectable until 90 min after exposure. In contrast, stimulation by anisomycin, a potent Jun-NH2-terminal kinase (JNK) agonist, exhibited no lag phase. An intermediate response was observed to emetine and cycloheximide, weak activators of JNK. 3. The potency of anisomycin to stimulate transport acutely (30 min of exposure) was 5- to 10-fold greater than for its chronic stimulation of transport, measured after 4 h of exposure. The stimulation of transport by a low concentration of anisomycin (0.3 microM) was transient, peaked at 30-60 min and it was inhibited (IC50 < 1 microM) by SB203580, which indicates that its mediator is not JNK, but the homologous p38(MAP kinase) (p38(MAPK)). In contrast, the responses to 4 h exposure to 300 microM anisomycin or puromycin were refractory to SB203580. 4. Exposure to anisomycin resulted in rapid activation of p38(MAPK). Activation of both p38(MAPK) and GLUT1 by 0.3 microM anisomycin was cancelled by puromycin. 5. We conclude that the activation of GLUT1 in response to anisomycin includes two components: a delayed component involving translational inhibition and a fast, puromycin-inhibitable component that is secondary to activation of p38(MAPK).
Barros
L F
LF
Department of Biochemistry and Molecular Biology, University of Leeds, UK. fbarros@bitmed.med.uchile.cl
Young
M
M
Saklatvala
J
J
Baldwin
S A
SA
D000273:Adipose Tissue / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D002478:Cells, Cultured; D002999:Clone Cells / Q000187:drug effects / Q000378:metabolism; D003847:Deoxyglucose / Q000378:metabolism; D004789:Enzyme Activation; D051272:Glucose Transporter Type 1; D007700:Kinetics; D020928:Mitogen-Activated Protein Kinases; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D013268:Stimulation, Chemical; D048051:p38 Mitogen-Activated Protein Kinases
D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; D011500:Protein Synthesis Inhibitors; D000841:Anisomycin; D003847:Deoxyglucose; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases