Characterization of the mitogen-activated protein kinase/90-kilodalton ribosomal protein S6 kinase signaling pathway in 3T3-L1 adipocytes and its role in insulin-stimulated glucose transport.
Insulin exerts diverse effects on mitogenesis, metabolism, gene expression, and protein synthesis depending on the target cell type. A variety of extracellular serine/threonine kinases, including the ribosomal protein S6 kinases pp70-ribosomal S6 kinase (pp70-S6K) and pp90-ribosomal S6 kinase (pp90rsk) and the erk-encoded mitogen-activated protein (MAP) kinases pp44mapk/ERK-1 and pp42mapk/ERK-2, have been postulated as mediators of insulin action. In this study, we have investigated the role of the MAP kinase/pp90rsk signaling pathway in insulin-stimulated glucose transport in 3T3-L1 adipocytes. Differentiation of 3T3-L1 fibroblasts into adipocyte-like cells was accompanied by a marked increase in the capacity of insulin to activate pp90rsk and pp44mapk. Whereas the maximal insulin-stimulated pp90rsk and pp44mapk activities were only approximately 30% of the serum-stimulated activities in preadipocytes, the insulin-stimulated kinase activities in adipocytes were equal to or greater than the serum-stimulated activities. The increase in hormone receptor number accompanying differentiation accounted for the greater sensitivity, as overexpression of human insulin receptors in NIH-3T3 cells also conferred insulin-stimulatable kinase activity. In 3T3-L1 adipocytes, the stimulation of pp90rsk and pp44mapk activities was sufficiently rapid and hormone sensitive to convey a signal for increased hexose uptake. However, epidermal growth factor and fetal bovine serum were equipotent with insulin in stimulating pp90rsk and pp44mapk activities in adipocytes, but were without effect on hexose uptake. These data indicate that activation of these enzymes is not sufficient for the acute stimulation of glucose transport.
Fingar
D C
DC
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115.
Birnbaum
M J
MJ
D016475:3T3 Cells; D017667:Adipocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D055504:Insulin Receptor Substrate Proteins; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D010750:Phosphoproteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D019893:Ribosomal Protein S6 Kinases; D015398:Signal Transduction; D014162:Transfection
C526292:IRS1 protein, human; D007328:Insulin; D055504:Insulin Receptor Substrate Proteins; C526295:Irs1 protein, mouse; D010750:Phosphoproteins; D011994:Recombinant Proteins; D017346:Protein Serine-Threonine Kinases; D019893:Ribosomal Protein S6 Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D005947:Glucose