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J Biol Chem, Vol. 274, Issue 15, 10071-10078, April 9, 1999

An Inhibitor of p38 Mitogen-activated Protein Kinase Prevents Insulin-stimulated Glucose Transport but Not Glucose Transporter Translocation in 3T3-L1 Adipocytes and L6 Myotubes

Gary SweeneyDagger , Romel SomwarDagger §, Toolsie RamlalDagger , Allen VolchukDagger §, Atsunori UeyamaDagger , and Amira KlipDagger §

From the Dagger  Programme in Cell Biology, Hospital for Sick Children and § Department of Biochemistry, University of Toronto, Toronto, Ontario, M5G 1X8 Canada

The precise mechanisms underlying insulin-stimulated glucose transport still require investigation. Here we assessed the effect of SB203580, an inhibitor of the p38 MAP kinase family, on insulin-stimulated glucose transport in 3T3-L1 adipocytes and L6 myotubes. We found that SB203580, but not its inactive analogue (SB202474), prevented insulin-stimulated glucose transport in both cell types with an IC50 similar to that for inhibition of p38 MAP kinase (0.6 µM). Basal glucose uptake was not affected. Moreover, SB203580 added only during the transport assay did not inhibit basal or insulin-stimulated transport. SB203580 did not inhibit insulin-stimulated translocation of the glucose transporters GLUT1 or GLUT4 in 3T3-L1 adipocytes as assessed by immunoblotting of subcellular fractions or by immunofluorescence of membrane lawns. L6 muscle cells expressing GLUT4 tagged on an extracellular domain with a Myc epitope (GLUT4myc) were used to assess the functional insertion of GLUT4 into the plasma membrane. SB203580 did not affect the insulin-induced gain in GLUT4myc exposure at the cell surface but largely reduced the stimulation of glucose uptake. SB203580 had no effect on insulin-dependent insulin receptor substrate-1 phosphorylation, association of the p85 subunit of phosphatidylinositol 3-kinase with insulin receptor substrate-1, nor on phosphatidylinositol 3-kinase, Akt1, Akt2, or Akt3 activities in 3T3-L1 adipocytes. In conclusion, in the presence of SB203580, insulin caused normal translocation and cell surface membrane insertion of glucose transporters without stimulating glucose transport. We propose that insulin stimulates two independent signals contributing to stimulation of glucose transport: phosphatidylinositol 3-kinase leads to glucose transporter translocation and a pathway involving p38 MAP kinase leads to activation of the recruited glucose transporter at the membrane.


Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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Protein Kinase B/Akt Mediates cAMP- and Cell Swelling-stimulated Na+/Taurocholate Cotransport and Ntcp Translocation
J. Biol. Chem., August 2, 2002; 277(32): 28578 - 28583.
[Abstract] [Full Text] [PDF]


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J. Appl. Physiol.Home page
K. Sakamoto and L. J. Goodyear
Exercise Effects on Muscle Insulin Signaling and Action: Invited Review: Intracellular signaling in contracting skeletal muscle
J Appl Physiol, July 1, 2002; 93(1): 369 - 383.
[Abstract] [Full Text] [PDF]


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CarcinogenesisHome page
J.-H. Cho, S.-D. Cho, H. Hu, S.-H. Kim, S. K. Lee, Y.-S. Lee, and K.-S. Kang
The roles of ERK1/2 and p38 MAP kinases in the preventive mechanisms of mushroom Phellinus linteus against the inhibition of gap junctional intercellular communication by hydrogen peroxide
Carcinogenesis, July 1, 2002; 23(7): 1163 - 1169.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
C. Huang, R. Somwar, N. Patel, W. Niu, D. Torok, and A. Klip
Sustained Exposure of L6 Myotubes to High Glucose and Insulin Decreases Insulin-Stimulated GLUT4 Translocation but Upregulates GLUT4 Activity
Diabetes, July 1, 2002; 51(7): 2090 - 2098.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
A. R. Gosmanov and D. B. Thomason
Insulin and Isoproterenol Differentially Regulate Mitogen-Activated Protein Kinase-Dependent Na+-K+-2Cl- Cotransporter Activity in Skeletal Muscle
Diabetes, March 1, 2002; 51(3): 615 - 623.
[Abstract] [Full Text] [PDF]


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BloodHome page
I. M. Pedersen, A. M. Buhl, P. Klausen, C. H. Geisler, and J. Jurlander
The chimeric anti-CD20 antibody rituximab induces apoptosis in B-cell chronic lymphocytic leukemia cells through a p38 mitogen activated protein-kinase-dependent mechanism
Blood, February 15, 2002; 99(4): 1314 - 1319.
[Abstract] [Full Text] [PDF]


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J. Appl. Physiol.Home page
C. Y. Christ, D. Hunt, J. Hancock, R. Garcia-Macedo, L. J. Mandarino, and J. L. Ivy
Exercise training improves muscle insulin resistance but not insulin receptor signaling in obese Zucker rats
J Appl Physiol, February 1, 2002; 92(2): 736 - 744.
[Abstract] [Full Text] [PDF]


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J. Clin. Endocrinol. Metab.Home page
X. Huang, A. Vaag, M. Hansson, and L. Groop
Down-Regulation of Insulin Receptor Substrates (IRS)-1 and IRS-2 and Src Homologous and Collagen-Like Protein Shc Gene Expression by Insulin in Skeletal Muscle Is Not Associated with Insulin Resistance or Type 2 Diabetes
J. Clin. Endocrinol. Metab., January 1, 2002; 87(1): 255 - 259.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
K. Lange
Role of microvillar cell surfaces in the regulation of glucose uptake and organization of energy metabolism
Am J Physiol Cell Physiol, January 1, 2002; 282(1): C1 - C26.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
A. R. Shikhman, D. C. Brinson, J. Valbracht, and M. K. Lotz
Cytokine Regulation of Facilitated Glucose Transport in Human Articular Chondrocytes
J. Immunol., December 15, 2001; 167(12): 7001 - 7008.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
R. Somwar, W. Niu, D. Y. Kim, G. Sweeney, V. K. Randhawa, C. Huang, T. Ramlal, and A. Klip
Differential Effects of Phosphatidylinositol 3-Kinase Inhibition on Intracellular Signals Regulating GLUT4 Translocation and Glucose Transport
J. Biol. Chem., November 30, 2001; 276(49): 46079 - 46087.
[Abstract] [Full Text] [PDF]


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J. Pharmacol. Exp. Ther.Home page
S. Apparsundaram, U. Sung, R. D. Price, and R. D. Blakely
Trafficking-Dependent and -Independent Pathways of Neurotransmitter Transporter Regulation Differentially Involving p38 Mitogen-Activated Protein Kinase Revealed in Studies of Insulin Modulation of Norepinephrine Transport in SK-N-SH Cells
J. Pharmacol. Exp. Ther., November 1, 2001; 299(2): 666 - 677.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
G. Sweeney, J. Keen, R. Somwar, D. Konrad, R. Garg, and A. Klip
High Leptin Levels Acutely Inhibit Insulin-Stimulated Glucose Uptake without Affecting Glucose Transporter 4 Translocation in L6 Rat Skeletal Muscle Cells
Endocrinology, November 1, 2001; 142(11): 4806 - 4812.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
F. Tremblay, C. Lavigne, H. Jacques, and A. Marette
Defective Insulin-Induced GLUT4 Translocation in Skeletal Muscle of High Fat-Fed Rats Is Associated With Alterations in Both Akt/Protein Kinase B and Atypical Protein Kinase C ({zeta}/{lambda}) Activities
Diabetes, August 1, 2001; 50(8): 1901 - 1910.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
N. Spravchikov, G. Sizyakov, M. Gartsbein, D. Accili, T. Tennenbaum, and E. Wertheimer
Glucose Effects on Skin Keratinocytes: Implications for Diabetes Skin Complications
Diabetes, July 1, 2001; 50(7): 1627 - 1635.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
D. Konrad, R. Somwar, G. Sweeney, K. Yaworsky, M. Hayashi, T. Ramlal, and A. Klip
The Antihyperglycemic Drug {alpha}-Lipoic Acid Stimulates Glucose Uptake via Both GLUT4 Translocation and GLUT4 Activation: Potential Role of p38 Mitogen-Activated Protein Kinase in GLUT4 Activation
Diabetes, June 1, 2001; 50(6): 1464 - 1471.
[Abstract] [Full Text]


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Am. J. Physiol. Cell Physiol.Home page
J.-Z. Zhang, W. Abbud, R. Prohaska, and F. Ismail-Beigi
Overexpression of stomatin depresses GLUT-1 glucose transporter activity
Am J Physiol Cell Physiol, May 1, 2001; 280(5): C1277 - C1283.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Heart Circ. Physiol.Home page
S. Schneider, W. Chen, J. Hou, C. Steenbergen, and E. Murphy
Inhibition of p38 MAPK {alpha}/{beta} reduces ischemic injury and does not block protective effects of preconditioning
Am J Physiol Heart Circ Physiol, February 1, 2001; 280(2): H499 - H508.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
G. Bandyopadhyay, Y. Kanoh, M. P. Sajan, M. L. Standaert, and R. V. Farese
Effects of Adenoviral Gene Transfer of Wild-Type, Constitutively Active, and Kinase-Defective Protein Kinase C-{lambda} on Insulin-Stimulated Glucose Transport in L6 Myotubes
Endocrinology, November 1, 2000; 141(11): 4120 - 4127.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Endocrinol. Metab.Home page
J. J. Wilkes and A. Bonen
Reduced insulin-stimulated glucose transport in denervated muscle is associated with impaired Akt-alpha activation
Am J Physiol Endocrinol Metab, October 1, 2000; 279(4): E912 - E919.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
G. Müller, S. Wied, and W. Frick
Cross Talk of pp125FAK and pp59Lyn Non-Receptor Tyrosine Kinases to Insulin-Mimetic Signaling in Adipocytes
Mol. Cell. Biol., July 1, 2000; 20(13): 4708 - 4723.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
H. Tong, W. Chen, R. E. London, E. Murphy, and C. Steenbergen
Preconditioning Enhanced Glucose Uptake Is Mediated by p38 MAP Kinase Not by Phosphatidylinositol 3-Kinase
J. Biol. Chem., April 14, 2000; 275(16): 11981 - 11986.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
F. V. Lali, A. E. Hunt, S. J. Turner, and B. M. J. Foxwell
The Pyridinyl Imidazole Inhibitor SB203580 Blocks Phosphoinositide-dependent Protein Kinase Activity, Protein Kinase B Phosphorylation, and Retinoblastoma Hyperphosphorylation in Interleukin-2-stimulated T Cells Independently of p38 Mitogen-activated Protein Kinase
J. Biol. Chem., March 15, 2000; 275(10): 7395 - 7402.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
A. S. Blair, E. Hajduch, G. J. Litherland, and H. S. Hundal
Regulation of Glucose Transport and Glycogen Synthesis in L6 Muscle Cells during Oxidative Stress. EVIDENCE FOR CROSS-TALK BETWEEN THE INSULIN AND SAPK2/p38 MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING PATHWAYS
J. Biol. Chem., December 17, 1999; 274(51): 36293 - 36299.
[Abstract] [Full Text] [PDF]


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Mol. Endocrinol.Home page
L. Braiman, A. Alt, T. Kuroki, M. Ohba, A. Bak, T. Tennenbaum, and S. R. Sampson
Protein Kinase C{delta} Mediates Insulin-Induced Glucose Transport in Primary Cultures of Rat Skeletal Muscle
Mol. Endocrinol., December 1, 1999; 13(12): 2002 - 2012.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
F. Roger, P.-Y. Martin, M. Rousselot, H. Favre, and E. Feraille
Cell Shrinkage Triggers the Activation of Mitogen-activated Protein Kinases by Hypertonicity in the Rat Kidney Medullary Thick Ascending Limb of the Henle's Loop. REQUIREMENT OF p38 KINASE FOR THE REGULATORY VOLUME INCREASE RESPONSE
J. Biol. Chem., November 26, 1999; 274(48): 34103 - 34110.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. F. Hausdorff, D. C. Fingar, K. Morioka, L. A. Garza, E. L. Whiteman, S. A. Summers, and M. J. Birnbaum
Identification of Wortmannin-sensitive Targets in 3T3-L1 Adipocytes. DISSOCIATION OF INSULIN-STIMULATED GLUCOSE UPTAKE AND GLUT4 TRANSLOCATION
J. Biol. Chem., August 27, 1999; 274(35): 24677 - 24684.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
R. C. Hresko and M. Mueckler
A Novel 68-kDa Adipocyte Protein Phosphorylated on Tyrosine in Response to Insulin and Osmotic Shock
J. Biol. Chem., June 9, 2000; 275(24): 18114 - 18120.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
Z. Huang, S.-L. Cheng, and E. Slatopolsky
Sustained Activation of the Extracellular Signal-regulated Kinase Pathway Is Required for Extracellular Calcium Stimulation of Human Osteoblast Proliferation
J. Biol. Chem., June 8, 2001; 276(24): 21351 - 21358.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
T. Suzuki, A. Hiroki, T. Watanabe, T. Yamashita, I. Takei, and K. Umezawa
Potentiation of Insulin-related Signal Transduction by a Novel Protein-tyrosine Phosphatase Inhibitor, Et-3,4-dephostatin, on Cultured 3T3-L1 Adipocytes
J. Biol. Chem., July 13, 2001; 276(29): 27511 - 27518.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M. Fujishiro, Y. Gotoh, H. Katagiri, H. Sakoda, T. Ogihara, M. Anai, Y. Onishi, H. Ono, M. Funaki, K. Inukai, et al.
MKK6/3 and p38 MAPK Pathway Activation Is Not Necessary for Insulin-induced Glucose Uptake but Regulates Glucose Transporter Expression
J. Biol. Chem., June 1, 2001; 276(23): 19800 - 19806.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
X. Xi, J. Han, and J.-Z. Zhang
Stimulation of Glucose Transport by AMP-activated Protein Kinase via Activation of p38 Mitogen-activated Protein Kinase
J. Biol. Chem., October 26, 2001; 276(44): 41029 - 41034.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
H. H. Versteeg, I. Hoedemaeker, S. H. Diks, J. C. Stam, M. Spaargaren, P. M. P. van Bergen en Henegouwen, S. J. H. van Deventer, and M. P. Peppelenbosch
Factor VIIa/Tissue Factor-induced Signaling via Activation of Src-like Kinases, Phosphatidylinositol 3-Kinase, and Rac
J. Biol. Chem., September 8, 2000; 275(37): 28750 - 28756.
[Abstract] [Full Text] [PDF]




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