The glucose-lowering agent sodium tungstate increases the levels and translocation of GLUT4 in L6 myotubes through a mechanism associated with ERK1/2 and MEF2D.
OBJECTIVE
The aim of this study was to investigate the action of the glucose-lowering compound sodium tungstate on glucose transport in muscle myotubes and to unravel the molecular events underlying the effects observed.


METHODS
We studied the effects of tungstate on 2-deoxy-D: -glucose uptake, levels and translocation of the glucose transporters GLUT4 and GLUT1, and Glut4 (also known as Slc2a4) promoter activity. We also measured the modifications of individual components of the signalling pathways involved in the effects observed.


RESULTS
Tungstate increased 2-deoxy-D: -glucose uptake in differentiated L6 myotubes through an increase in the total amount and translocation of GLUT4 transporter. The effects on glucose uptake were additive to those of insulin. Tungstate activated transcription of the Glut4 promoter, as shown by an increase in Glut4 mRNA, and by a promoter reporter assay. The assay of deletions of the Glut4 promoter indicated that the effect of tungstate is mediated by the myocyte enhancer factor 2 (MEF2)-binding domain. Accordingly, MEF2 levels and DNA binding activities were increased in response to the treatment. Tungstate-induced glucose uptake and GLUT4 transcriptional activation were dependent on the activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), while no changes were observed in the phosphorylation state of the beta subunit of the insulin receptor, in the phosphatidylinositol 3-kinase pathway or in the activation of 5'AMP-activated protein kinase.


CONCLUSIONS
Tungstate activates glucose uptake in myotubes through a novel ERK1/2-dependent mechanism. This effect is exerted by an increase in the content and translocation of the GLUT4 transporter. This is the first report of a glucose-lowering compound activating Glut4 transcription through an ERK1/2-dependent increase in MEF2 levels.
Girón
M D
MD
Department of Biochemistry and Molecular Biology II, School of Pharmacy, University of Granada, Campus de Cartuja sn, 18071, Granada, Spain.
Sevillano
N
N
Vargas
A M
AM
Domínguez
J
J
Guinovart
J J
JJ
Salto
R
R
D000818:Animals; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000378:metabolism / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D051275:Glucose Transporter Type 4 / Q000235:genetics* / Q000378:metabolism*; D006943:Hyperglycemia / Q000188:drug therapy / Q000503:physiopathology; D020935:MAP Kinase Signaling System / Q000187:drug effects* / Q000502:physiology; D064326:MEF2 Transcription Factors; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D018006:Myogenic Regulatory Factors / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014316:Tritium; D017973:Tungsten Compounds / Q000494:pharmacology*
D051275:Glucose Transporter Type 4; D064326:MEF2 Transcription Factors; D018006:Myogenic Regulatory Factors; C495694:Slc2a4 protein, rat; D017973:Tungsten Compounds; D014316:Tritium; C025399:sodium tungstate(VI); D003847:Deoxyglucose; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D005947:Glucose