Interleukin-3-mediated cell survival signals include phosphatidylinositol 3-kinase-dependent translocation of the glucose transporter GLUT1 to the cell surface.
Maintenance of glucose uptake is a key component in the response of hematopoietic cells to survival factors. To investigate the mechanism of this response we employed the interleukin-3 (IL-3)-dependent murine mast cell line IC2.9. In these cells, hexose uptake decreased markedly upon withdrawal of IL-3, whereas its readdition led to rapid (t(1/2) approximately 10 min) stimulation of transport, associated with an approximately 4-fold increase in Vmax but no change in Km. Immunocytochemistry and photoaffinity labeling revealed that IL-3 caused translocation of intracellular GLUT1 transporters to the cell surface, whereas a second transporter isoform, GLUT3, remained predominantly intracellular. The inhibitory effects of latrunculin B and jasplakinolide, and of nocodazole and colchicine, respectively, revealed a requirement for both the actin and microtubule cytoskeletons in GLUT1 translocation and transport stimulation. Both IL-3 stimulation of transport and GLUT1 translocation were also prevented by the phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002. The time courses for activation of phosphatidylinositol 3-kinase and its downstream target, protein kinase B, by IL-3 were consistent with a role in IL-3-induced transporter translocation and enhanced glucose uptake. We conclude that one component of the survival mechanisms elicited by IL-3 involves the subcellular redistribution of glucose transporters, thus ensuring the supply of a key metabolic substrate.
Bentley
Johanne
J
School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom
Itchayanan
Dalina
D
Barnes
Kay
K
McIntosh
Elizabeth
E
Tang
Xiuwen
X
Downes
C Peter
CP
Holman
Geoffrey D
GD
Whetton
Anthony D
AD
Owen-Lynch
P Jane
PJ
Baldwin
Stephen A
SA
D000199:Actins / Q000378:metabolism; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001693:Biological Transport, Active; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D003847:Deoxyglucose / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D051272:Glucose Transporter Type 1; D051274:Glucose Transporter Type 3; D007377:Interleukin-3 / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D009419:Nerve Tissue Proteins; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction; D000077191:Wortmannin
D000199:Actins; D000730:Androstadienes; D002867:Chromones; D003847:Deoxyglucose; D004791:Enzyme Inhibitors; D005947:Glucose; D051272:Glucose Transporter Type 1; D051274:Glucose Transporter Type 3; D007377:Interleukin-3; D009004:Monosaccharide Transport Proteins; D009025:Morpholines; D009419:Nerve Tissue Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D011994:Recombinant Proteins; D000077191:Wortmannin; C495678:Slc2a1 protein, mouse; C495689:Slc2a3 protein, mouse; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one