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Second Department of Internal Medicine (H.S., W.O., M.T., S.K.,
K.K., M.M., M.S., M.K.), Kobe University School of Medicine, Kobe
650, Japan,
Molecular Cardiology Unit (S.N., H.U.), Kyushu
University School of Medicine, Fukuoka 812-82, Japan
The regulatory mechanism of glucose uptake in
3T3-L1 adipocytes was investigated with the use of recombinant
adenovirus vectors encoding various dominant negative proteins.
Infection with a virus encoding a mutant regulatory subunit of
phosphoinositide (PI) 3-kinase that does not bind the 110-kDa
catalytic subunit (
p85) inhibited the insulin-induced increase
in PI 3-kinase activity coprecipitated by antibodies to phosphotyrosine
and glucose uptake in a virus dose-dependent manner. Overexpression of
a dominant negative RAS mutant in which Asp57
is replaced with tyrosine (RAS57Y) or of a dominant negative SOS mutant
that lacks guanine nucleotide exchange activity (SOS) abolished the
insulin-induced increase in mitogen-activated protein kinase activity,
but had no effect on PI 3-kinase activity or glucose uptake. Although
GH and hyperosmolarity attributable to 300 mM
sorbitol each promoted glucose uptake and translocation of glucose
transporter (GLUT)4 to an extent comparable to that of insulin, these
stimuli triggered little or no association of PI 3-kinase activity with
tyrosine-phosphorylated proteins. Overexpression of p85 or treatment
of cells with wortmannin, an inhibitor of PI 3-kinase activity, had no
effect on glucose uptake or translocation of GLUT4 stimulated by GH or
hyperosmolarity. Moreover, overexpression of SOS or RAC17N also did
not affect the increase in glucose uptake induced by these stimuli. A
serine/threonine kinase Akt, a constitutively active mutant of which
was previously shown to stimulate glucose uptake, is activated by
insulin, GH, and hyperosmolarity to 
4-fold, 2.1-fold, and
2.3-fold over basal level, respectively. These results suggest that
insulin-induced but neither GH- or hyperosmolarity-induced glucose
uptake is PI 3-kinase-dependent, and neither RAS nor RAC is required
for glucose uptake induced by these stimuli in 3T3-L1 adipocytes.
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K. Kotani, W. Ogawa, Y. Hino, T. Kitamura, H. Ueno, W. Sano, C. Sutherland, D. K. Granner, and M. Kasuga Dominant Negative Forms of Akt (Protein Kinase B) and Atypical Protein Kinase Clambda Do Not Prevent Insulin Inhibition of Phosphoenolpyruvate Carboxykinase Gene Transcription J. Biol. Chem., July 23, 1999; 274(30): 21305 - 21312. [Abstract] [Full Text] [PDF]
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M. Takata, W. Ogawa, T. Kitamura, Y. Hino, S. Kuroda, K. Kotani, A. Klip, A.-C. Gingras, N. Sonenberg, and M. Kasuga Requirement for Akt (Protein Kinase B) in Insulin-induced Activation of Glycogen Synthase and Phosphorylation of 4E-BP1 (PHAS-1) J. Biol. Chem., July 16, 1999; 274(29): 20611 - 20618. [Abstract] [Full Text] [PDF]
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S. Takasuga, T. Katada, M. Ui, and O. Hazeki Enhancement by Adenosine of Insulin-induced Activation of Phosphoinositide 3-Kinase and Protein Kinase B in Rat Adipocytes J. Biol. Chem., July 9, 1999; 274(28): 19545 - 19550. [Abstract] [Full Text] [PDF]
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D. Chen, R. V. Fucini, A. L. Olson, B. A. Hemmings, and J. E. Pessin Osmotic Shock Inhibits Insulin Signaling by Maintaining Akt/Protein Kinase B in an Inactive Dephosphorylated State Mol. Cell. Biol., July 1, 1999; 19(7): 4684 - 4694. [Abstract] [Full Text] [PDF]
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D. J. Sherwood, S. D. Dufresne, J. F. Markuns, B. Cheatham, D. E. Moller, D. Aronson, and L. J. Goodyear Differential regulation of MAP kinase, p70S6K, and Akt by contraction and insulin in rat skeletal muscle Am J Physiol Endocrinol Metab, May 1, 1999; 276(5): E870 - E878. [Abstract] [Full Text] [PDF]
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A. C. P. Thirone, C. R. O. Carvalho, and M. J. A. Saad Growth Hormone Stimulates the Tyrosine Kinase Activity of JAK2 and Induces Tyrosine Phosphorylation of Insulin Receptor Substrates and Shc in Rat Tissues Endocrinology, January 1, 1999; 140(1): 55 - 62. [Abstract] [Full Text]
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K. Kotani, W. Ogawa, M. Matsumoto, T. Kitamura, H. Sakaue, Y. Hino, K. Miyake, W. Sano, K. Akimoto, S. Ohno, et al. Requirement of Atypical Protein Kinase Clambda for Insulin Stimulation of Glucose Uptake but Not for Akt Activation in 3T3-L1 Adipocytes Mol. Cell. Biol., December 1, 1998; 18(12): 6971 - 6982. [Abstract] [Full Text]
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H. Sakaue, W. Ogawa, M. Matsumoto, S. Kuroda, M. Takata, T. Sugimoto, B. M. Spiegelman, and M. Kasuga Posttranscriptional Control of Adipocyte Differentiation through Activation of Phosphoinositide 3-Kinase J. Biol. Chem., October 30, 1998; 273(44): 28945 - 28952. [Abstract] [Full Text] [PDF]
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P. A. Staubs, J. G. Nelson, D. R. Reichart, and J. M. Olefsky Platelet-derived Growth Factor Inhibits Insulin Stimulation of Insulin Receptor Substrate-1-associated Phosphatidylinositol 3-Kinase in 3T3-L1 Adipocytes without Affecting Glucose Transport J. Biol. Chem., September 25, 1998; 273(39): 25139 - 25147. [Abstract] [Full Text] [PDF]
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D. Wang and H. S. Sul Insulin Stimulation of the Fatty Acid Synthase Promoter Is Mediated by the Phosphatidylinositol 3-Kinase Pathway. INVOLVEMENT OF PROTEIN KINASE B/Akt J. Biol. Chem., September 25, 1998; 273(39): 25420 - 25426. [Abstract] [Full Text] [PDF]
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Y. Tamori, M. Kawanishi, T. Niki, H. Shinoda, S. Araki, H. Okazawa, and M. Kasuga Inhibition of Insulin-induced GLUT4 Translocation by Munc18c through Interaction with Syntaxin4 in 3T3-L1 Adipocytes J. Biol. Chem., July 31, 1998; 273(31): 19740 - 19746. [Abstract] [Full Text] [PDF]
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T. Kitamura, W. Ogawa, H. Sakaue, Y. Hino, S. Kuroda, M. Takata, M. Matsumoto, T. Maeda, H. Konishi, U. Kikkawa, et al. Requirement for Activation of the Serine-Threonine Kinase Akt (Protein Kinase B) in Insulin Stimulation of Protein Synthesis but Not of Glucose Transport Mol. Cell. Biol., July 1, 1998; 18(7): 3708 - 3717. [Abstract] [Full Text]
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T. Yamauchi, Y. Kaburagi, K. Ueki, Y. Tsuji, G. R. Stark, I. M. Kerr, T. Tsushima, Y. Akanuma, I. Komuro, K. Tobe, et al. Growth Hormone and Prolactin Stimulate Tyrosine Phosphorylation of Insulin Receptor Substrate-1, -2, and -3, Their Association with p85 Phosphatidylinositol 3-Kinase (PI3-kinase), and Concomitantly PI3-kinase Activation via JAK2 Kinase J. Biol. Chem., June 19, 1998; 273(25): 15719 - 15726. [Abstract] [Full Text] [PDF]
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J. T. Brozinick Jr. and M. J. Birnbaum Insulin, but Not Contraction, Activates Akt/PKB in Isolated Rat Skeletal Muscle J. Biol. Chem., June 12, 1998; 273(24): 14679 - 14682. [Abstract] [Full Text] [PDF]
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A. Janez, D. S. Worrall, T. Imamura, P. M. Sharma, and J. M. Olefsky The Osmotic Shock-induced Glucose Transport Pathway in 3T3-L1 Adipocytes Is Mediated by Gab-1 and Requires Gab-1-associated Phosphatidylinositol 3-Kinase Activity for Full Activation J. Biol. Chem., August 25, 2000; 275(35): 26870 - 26876. [Abstract] [Full Text] [PDF]
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K. Kotani, W. Ogawa, M. Hashiramoto, T. Onishi, S. Ohno, and M. Kasuga Inhibition of Insulin-induced Glucose Uptake by Atypical Protein Kinase C Isotype-specific Interacting Protein in 3T3-L1 Adipocytes J. Biol. Chem., August 18, 2000; 275(34): 26390 - 26395. [Abstract] [Full Text] [PDF]
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A. V. Bakin, A. K. Tomlinson, N. A. Bhowmick, H. L. Moses, and C. L. Arteaga Phosphatidylinositol 3-Kinase Function Is Required for Transforming Growth Factor beta -mediated Epithelial to Mesenchymal Transition and Cell Migration J. Biol. Chem., November 17, 2000; 275(47): 36803 - 36810. [Abstract] [Full Text] [PDF]
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M. Matsumoto, W. Ogawa, Y. Hino, K. Furukawa, Y. Ono, M. Takahashi, M. Ohba, T. Kuroki, and M. Kasuga Inhibition of Insulin-induced Activation of Akt by a Kinase-deficient Mutant of the epsilon Isozyme of Protein Kinase C J. Biol. Chem., April 20, 2001; 276(17): 14400 - 14406. [Abstract] [Full Text] [PDF]
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K. Nakajima, K. Yamauchi, S. Shigematsu, S. Ikeo, M. Komatsu, T. Aizawa, and K. Hashizume Selective Attenuation of Metabolic Branch of Insulin Receptor Down-signaling by High Glucose in a Hepatoma Cell Line, HepG2 Cells J. Biol. Chem., June 30, 2000; 275(27): 20880 - 20886. [Abstract] [Full Text] [PDF]
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