Myocyte enhancer factor 2 (MEF2) binding site is essential for C2C12 myotube-specific expression of the rat GLUT4/muscle-adipose facilitative glucose transporter gene

Min-Ling Liu; Ann Louise Olson; Nicholas P Edgington; W. Scott Moye‐Rowley; Jeffrey E Pessin

doi:10.1016/S0021-9258(18)46957-8

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Myocyte enhancer factor 2 (MEF2) binding site is essential for C2C12 myotube-specific expression of the rat GLUT4/muscle-adipose facilitative glucose transporter gene

Journal article

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Peer reviewed

Myocyte enhancer factor 2 (MEF2) binding site is essential for C2C12 myotube-specific expression of the rat GLUT4/muscle-adipose facilitative glucose transporter gene

Min-Ling Liu, Ann Louise Olson, Nicholas P Edgington, W. Scott Moye‐Rowley and Jeffrey E Pessin

The Journal of biological chemistry, Vol.269(45), pp.28514-28521

11/11/1994

DOI: 10.1016/S0021-9258(18)46957-8

PMID: 7545962

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Abstract

We have cloned and characterized the rat GLUT4 gene in order to identify the cis-DNA elements responsible for tissue-specific GLUT4 expression. In this study, a variety of luciferase reporter gene constructs were transiently transfected into C2C12 myoblasts and myotubes as a model for skeletal muscle differentiation. These data identified a 103-base pair fragment, located from -522 to -420 relative to the transcription initiation site, that was sufficient to account for GLUT4 C2C12 myotube-specific expression. This fragment was operationally defined as an enhancer since it conferred myotube-specific expression in the context of both the minimal native GLUT4 or the heterologous thymidine kinase promoters in an orientation-independent manner. Further, mutagenesis of this fragment demonstrated that a sequence analogous to the muscle creatine kinase myocyte enhancer factor 2 (MEF2) binding site (-466 and -457) was required for transcriptional activation. Electrophoretic mobility gel shift assays demonstrated specific binding activity to the GLUT4 MEF2 sequences which directly correlated with functional expression. Although this element was capable of directing myotube-specific expression when cloned as multiple copies into luciferase reporter gene constructs, the MEF2 sequence alone was insufficient to enhance GLUT4 expression. These data demonstrated that GLUT4 muscle-specific expression is conferred by a 103-base pair DNA sequence located between -522 and -420 of rat GLUT4 gene. This region encompasses a MEF2 binding site which was necessary, but not sufficient, for transcriptional activation.

Cell Differentiation

Gene Expression

Transfection

Adipose Tissue - metabolism

Animals

Base Sequence

Binding Sites

Cell Nucleus - metabolism

Cells, Cultured

Consensus Sequence

Creatine Kinase - genetics

DNA - chemistry

DNA - metabolism

DNA Primers

DNA-Binding Proteins - metabolism

Enhancer Elements, Genetic

Glucose Transporter Type 4

HeLa Cells

Humans

Luciferases - biosynthesis

MEF2 Transcription Factors

Molecular Sequence Data

Monosaccharide Transport Proteins - biosynthesis

Muscle Proteins

Muscles - cytology

Muscles - metabolism

Myocardium - metabolism

Myogenic Regulatory Factors

Rats

Recombinant Proteins - biosynthesis

Sequence Deletion

Thymidine Kinase - biosynthesis

Thymidine Kinase - genetics

Transcription Factors - metabolism

Details

Title: Subtitle: Myocyte enhancer factor 2 (MEF2) binding site is essential for C2C12 myotube-specific expression of the rat GLUT4/muscle-adipose facilitative glucose transporter gene
Creators: Min-Ling Liu - University of Iowa
Ann Louise Olson - University of Iowa
Nicholas P Edgington - University of Iowa
W. Scott Moye‐Rowley - University of Iowa
Jeffrey E Pessin - University of Iowa
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.269(45), pp.28514-28521
DOI: 10.1016/S0021-9258(18)46957-8
PMID: 7545962
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Grant note: DK42452 / NIDDK NIH HHS DK25925 / NIDDK NIH HHS DK44612 / NIDDK NIH HHS
Language: English
Date published: 11/11/1994
Academic Unit: Molecular Physiology and Biophysics; Computer Science; Internal Medicine
Record Identifier: 9984297501002771

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