title: Ectopic expression of protein kinase CbetaII, -delta, and -epsilon, but not -betaI or -zeta, provide for insulin stimulation of glucose uptake in NIH-3T3 cells.
abstract: Insulin regulates a diverse array of signaling pathways involved in the control of growth, differentiation, proliferation, and metabolism. Insulin increases in glucose uptake via a protein kinase C-dependent pathway in target tissues such as fat and muscle are well documented. Insulin-regulated events, however, occur in all cells. The utilization of glucose as a preferred energy source is a ubiquitous event in eukaryotic cells. In NIH-3T3 fibroblasts, insulin treatment increased levels of the cPKC and nPKC activator, diacylglycerol. Insulin-responsive 2-[(3)H]deoxyglucose uptake was stimulated in a dose-dependent manner. The overexpression of protein kinase C (PKC)betaI, -betaII, -delta, -epsilon, and -zeta was used to investigate the specificity of PKC isozymes for insulin-sensitive glucose uptake. The stable overexpression of PKCbetaII, -delta, and -epsilon resulted in increases in insulin-stimulated 2-[(3)H]deoxyglucose uptake compared to vector control cells, while basal 2-deoxyglucose uptake levels were not elevated. Overexpression of PKCbetaI and PKCzeta isozymes had no further effect on basal or insulin-stimulated 2-deoxyglucose uptake. The PKC-specific inhibitor, CGP41251, blocked insulin effects on 2-deoxyglucose uptake but not its effects on tyrosine phosphorylation of cellular substrates. Insulin-stimulated 3-O-methylglucose uptake was also greater in cells overexpressing PKCbetaII, -delta, and -epsilon, compared to control cells. The increased responsiveness was not accompanied by conversion of 3T3 cells to the adipocyte phenotype or the increased expression of insulin receptors or glucose transporters (GLUT1-type). Insulin-stimulated recruitment of GLUT1 to plasma membranes of cells overexpressing PKCbetaII, -delta, and -epsilon, was greater than that in control cells. The data suggest that more than one PKC isozyme is involved in insulin signaling pathways in fibroblasts, resulting in increased GLUT1 transporter recruitment to cell membranes.
authors.affiliation: James A. Haley Veterans Hospital, University of South Florida College of Medicine, Tampa, Florida, 33612, USA. dcooper@com1.med.usf.edu
authors.forename: D R
authors.initials: DR
authors.lastname: Cooper
authors.forename: J E
authors.initials: JE
authors.lastname: Watson
authors.forename: N
authors.initials: N
authors.lastname: Patel
authors.forename: P
authors.initials: P
authors.lastname: Illingworth
authors.forename: M
authors.initials: M
authors.lastname: Acevedo-Duncan
authors.forename: J
authors.initials: J
authors.lastname: Goodnight
authors.forename: C E
authors.initials: CE
authors.lastname: Chalfant
authors.forename: H
authors.initials: H
authors.lastname: Mischak
mesh_terms: D019325:3-O-Methylglucose / Q000378:metabolism; D016475:3T3 Cells; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D003847:Deoxyglucose / Q000378:metabolism; D004075:Diglycerides / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1; D007328:Insulin / Q000494:pharmacology*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011972:Receptor, Insulin / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D019311:Staurosporine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D004075:Diglycerides; D004791:Enzyme Inhibitors; D051272:Glucose Transporter Type 1; D007328:Insulin; D007527:Isoenzymes; D009004:Monosaccharide Transport Proteins; D011994:Recombinant Proteins; C495678:Slc2a1 protein, mouse; D019325:3-O-Methylglucose; D003847:Deoxyglucose; D011972:Receptor, Insulin; D011493:Protein Kinase C; D019311:Staurosporine; C059539:midostaurin; D005947:Glucose