Akt mediates insulin induction of glucose uptake and up‐regulation of GLUT4 gene expression in brown adipocytes

FEBS Letters

Lorenzo, Margarita¹  Hernandez, Rosario¹  Teruel, Teresa¹ 

[1] Departamento de Bioquimica y Biologia Molecular II, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain

关键词: Glucose uptake;    GLUT4 translocation;    GLUT4 gene expression;    Akt;    Insulin;    BSA;    bovine serum albumin;    MEM;    minimal essential medium;    PBS;    phosphate-buffered saline;    ECL;    enhanced chemiluminescence;    FCS;    foetal calf serum;    PK;    protein kinase;    PI;    phosphatidylinositol;    PMSF;    phenylmethylsulfonyl fluoride;    CAT;    chloramphenicol acetyltransferase;    GFP;    green fluorescent protein;    gal;    galactosidase;   

DOI  :  10.1016/S0014-5793(01)02353-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Insulin acutely stimulated glucose uptake in rat primary brown adipocytes in a PI3-kinase-dependent but p70S6-kinase-independent manner. Since Akt represents an intermediate step between these kinases, this study investigated the contribution of Akt to insulin-induced glucose uptake by the use of a chemical compound, ML-9, as well as by transfection with a dominant-negative form of Akt (ΔAkt). Pretreatment with ML-9 for 10 min completely inhibited insulin stimulation of (1) Akt kinase activity, (2) Akt phosphorylation on the regulatory residue Ser473 but not on Thr308, and (3) mobility shift in Akt1 and Akt2. However, ML-9 did not affect insulin-stimulated PI3-kinase nor PKCζ activities. In consequence, ML-9 precluded insulin stimulation of glucose uptake and GLUT4 translocation to plasma membrane (determined by Western blot), without any effect on the basal glucose uptake. Moreover, ΔAkt impaired insulin stimulation of glucose uptake and GFP-tagged GLUT4 translocation to plasma membrane in transiently transfected immortalised brown adipocytes and HeLa cells, respectively. Furthermore, ML-9 treatment for 6 h down-regulated insulin-induced GLUT4 mRNA accumulation, without affecting GLUT1 expression, in a similar fashion as LY294002. Indeed, co-transfection of brown adipocytes with ΔAkt precluded the transactivation of GLUT4-CAT promoter by insulin in a similar fashion as a dominant-negative form of PI3-kinase. Our results indicate that activation of Akt may be an essential requirement for insulin regulation of glucose uptake and GLUT4 gene expression in brown adipocytes.

【 授权许可】

Unknown