IGF-1 controls GLUT3 expression in muscle via the transcriptional factor Sp1

John A. Copland, Aaron W. Pardini, Thomas Wood, Deling Yin, Allan Green, Yvonne H. Bodenburg, Randall Urban, Charles A. Stuart

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Glucose transporter 3 (GLUT3), while first found in human fetal muscle, is predominantly expressed in brain and neural tissue. By several independent techniques we have previously shown that GLUT3 is expressed in human skeletal muscle cells. The structure of the human GLUT3 gene has not been previously reported nor has there been any evaluation of the 5′-untranslated region (UTR). To this end, we have cloned and sequenced the human GLUT3 gene. Insulin-like growth factor-1 (IGF-1) increased endogenous Glut3 protein in cultured L6 myotubes, and similarly stimulated luciferase activity in a construct of the human GLUT3 5′-UTR linked to a luciferase reporter gene. Actinomycin D, an inhibitor of mRNA synthesis, prevented IGF-1 stimulation of Glut3 protein. Transfection of L6 cells with Sp1 increased Glut3 and augmented IGF-1 stimulation of Glut3 expression. Knockdown of Glut3 expression in cultured L6 muscle cells using small interference RNA (siRNA) specific for Glut3 significantly reduced myocyte glucose uptake. DNAse footprinting and gel shift assays showed Sp1 specifically bound to the human GLUT3 5′-UTR. Substitution mutants of the human GLUT3 5′-UTR luciferase construct indicated that only one of three Sp1 site clusters was involved in IGF-1 action. These data, using both a human GLUT3 5′-UTR construct and L6 cells' endogenous promoter, suggest that IGF-1 plays a role in maintaining muscle GLUT3 expression and basal glucose uptake via the transcriptional factor Sp1.

Original languageEnglish (US)
Pages (from-to)631-640
Number of pages10
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1769
Issue number11-12
DOIs
StatePublished - Nov 2007

Fingerprint

Facilitative Glucose Transport Proteins
Somatomedins
Muscle
5' Untranslated Regions
Muscles
3' Untranslated Regions
Luciferases
Muscle Cells
Genes
Cells
Glucose
Skeletal Muscle Fibers
Dactinomycin
RNA Interference
Reporter Genes
Transfection
Assays
Brain
Skeletal Muscle
Proteins

Keywords

  • GLUT3
  • IGF-1
  • Muscle
  • Sp1

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology
  • Biophysics

Cite this

Copland, J. A., Pardini, A. W., Wood, T., Yin, D., Green, A., Bodenburg, Y. H., ... Stuart, C. A. (2007). IGF-1 controls GLUT3 expression in muscle via the transcriptional factor Sp1. Biochimica et Biophysica Acta - Gene Structure and Expression, 1769(11-12), 631-640. https://doi.org/10.1016/j.bbaexp.2007.08.002

IGF-1 controls GLUT3 expression in muscle via the transcriptional factor Sp1. / Copland, John A.; Pardini, Aaron W.; Wood, Thomas; Yin, Deling; Green, Allan; Bodenburg, Yvonne H.; Urban, Randall; Stuart, Charles A.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1769, No. 11-12, 11.2007, p. 631-640.

Research output: Contribution to journalArticle

Copland, John A. ; Pardini, Aaron W. ; Wood, Thomas ; Yin, Deling ; Green, Allan ; Bodenburg, Yvonne H. ; Urban, Randall ; Stuart, Charles A. / IGF-1 controls GLUT3 expression in muscle via the transcriptional factor Sp1. In: Biochimica et Biophysica Acta - Gene Structure and Expression. 2007 ; Vol. 1769, No. 11-12. pp. 631-640.
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AB - Glucose transporter 3 (GLUT3), while first found in human fetal muscle, is predominantly expressed in brain and neural tissue. By several independent techniques we have previously shown that GLUT3 is expressed in human skeletal muscle cells. The structure of the human GLUT3 gene has not been previously reported nor has there been any evaluation of the 5′-untranslated region (UTR). To this end, we have cloned and sequenced the human GLUT3 gene. Insulin-like growth factor-1 (IGF-1) increased endogenous Glut3 protein in cultured L6 myotubes, and similarly stimulated luciferase activity in a construct of the human GLUT3 5′-UTR linked to a luciferase reporter gene. Actinomycin D, an inhibitor of mRNA synthesis, prevented IGF-1 stimulation of Glut3 protein. Transfection of L6 cells with Sp1 increased Glut3 and augmented IGF-1 stimulation of Glut3 expression. Knockdown of Glut3 expression in cultured L6 muscle cells using small interference RNA (siRNA) specific for Glut3 significantly reduced myocyte glucose uptake. DNAse footprinting and gel shift assays showed Sp1 specifically bound to the human GLUT3 5′-UTR. Substitution mutants of the human GLUT3 5′-UTR luciferase construct indicated that only one of three Sp1 site clusters was involved in IGF-1 action. These data, using both a human GLUT3 5′-UTR construct and L6 cells' endogenous promoter, suggest that IGF-1 plays a role in maintaining muscle GLUT3 expression and basal glucose uptake via the transcriptional factor Sp1.

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