Interaction of urokinase-type plasminogen activator with its receptor rapidly induces activation of glucose transporters.
The interaction of urokinase-type plasminogen activator (u-PA) or of u-PA amino-terminal fragment (u-PA-ATF) with the cell surface receptor (u-PAR) was found to stimulate an increase of glucose uptake in many cell lines, ranging from normal and transformed human fibroblasts, mouse fibroblasts transfected with human u-PAR, and cells of epidermal origin. Such increase of glucose uptake reached a peak within 5-10 min, depending on the cell line, and occurred through the facilitative glucose transporters (GLUTs), since it was inhibited by cytochalasin B. Each cell line showed a specific mosaic of glucose transporter isoforms, GLUT2 being the most widespread and GLUT1 the most abundant, when present. u-PAR stimulation was followed by translocation of GLUT1 from the microsomal to the membrane compartment, as shown by both immunoblotting and immunofluorescence of sonicated plasma membrane sheets and by activation of GLUT2 on the cell surface. Both translocation and activation resulted inhibitable by protein-tyrosine kinase inhibitors and independent of downregulation of protein kinase C (PKC). The increase of intracellular glucose was followed by neosynthesis of diacylglycerol (DAG) from glucose, as previously shown. Such neosynthesis was completely inhibited by impairment of facilitative GLUT transport by cytochalasin B. DAG neosynthesis was followed by activation of PKC, whose activity translocated into the intracellular compartment (PKM), where it probably phosphorylates substrates required for u-PAR-dependent chemotaxis. Our data show that u-PAR-mediated signal transduction, related with u-PA-induced chemotaxis, involves activation of tyrosine kinase-dependent glucose transporters, leading to increased de novo DAG synthesis from glucose, eventually resulting in activation of PKC.
Anichini
E
E
Istituto di Patologia Generale, Universita di Firenze, Italy.
Zamperini
A
A
Chevanne
M
M
Caldini
R
R
Pucci
M
M
Fibbi
G
G
Del Rosso
M
M
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism; D003571:Cytochalasin B / Q000494:pharmacology; D003847:Deoxyglucose / Q000378:metabolism*; D004075:Diglycerides / Q000096:biosynthesis; D005347:Fibroblasts; D051272:Glucose Transporter Type 1; D051273:Glucose Transporter Type 2; D006801:Humans; D007700:Kinetics; D008168:Lung; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis*; D010446:Peptide Fragments / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000096:biosynthesis; D013539:Simian virus 40; D012867:Skin; D013347:Subcellular Fractions / Q000201:enzymology; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D004075:Diglycerides; D051272:Glucose Transporter Type 1; D051273:Glucose Transporter Type 2; D009004:Monosaccharide Transport Proteins; C524898:PLAUR protein, human; D010446:Peptide Fragments; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; C495677:SLC2A1 protein, human; C495678:Slc2a1 protein, mouse; D003571:Cytochalasin B; D003847:Deoxyglucose; D011493:Protein Kinase C; D014568:Urokinase-Type Plasminogen Activator