Identification of a skeletal muscle-specific regulatory domain in the rat GLUT4/muscle-fat gene

Jeanne M. Richardson, Jeffrey E. Pessin

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

To identify sequences responsible for the muscle-specific expression of the rat GLUT4/muscle-fat gene, we examined the transcriptional regulation of this gene in the differentiating murine C2C12 skeletal muscle cell line. Differentiated myofibers displayed a 4-5-fold increase in GLUT4 mRNA compared with undifferentiated myoblasts which paralleled the conversion from non-muscle β-actin mRNA to muscle-specific α-actin mRNA expression. Transient transfection of progressive 5′ and 3′ deletions of the GLUT4 5′-flanking DNA identified a 281-base pair region located between -517 and -237 relative to the transcription start site which conferred myotube-specific expression. This region increased reporter activity in the context of the GLUT4 minimal promoter in an orientation-independent manner and, in addition, onto the heterologous thymidine kinase promoter. Myotube-specific expression of both GLUT4 reporter constructs and the endogenous mouse GLUT4 mRNA was also observed to be thyroid hormone-dependent. Further, cotransfection of reporter constructs containing the 281-base pair GLUT4 differentiation-specific enhancer with the thyroid hormone receptor specifically increased luciferase activity in myotubes approximately 12-fold. Thus, these data demonstrate the presence of a proximal skeletal muscle-specific activation domain that is necessary for both myotube-specific GLUT4 expression and thyroid hormone responsiveness.

Original languageEnglish (US)
Pages (from-to)21021-21027
Number of pages7
JournalJournal of Biological Chemistry
Volume268
Issue number28
StatePublished - Oct 5 1993
Externally publishedYes

Fingerprint

Skeletal Muscle Fibers
Muscle
Rats
Skeletal Muscle
Genes
Fats
Muscles
Messenger RNA
Thyroid Hormones
Base Pairing
Actins
Thyroid Hormone Receptors
Thymidine Kinase
Myoblasts
Transcription Initiation Site
Luciferases
Muscle Cells
Transfection
Cell Line
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of a skeletal muscle-specific regulatory domain in the rat GLUT4/muscle-fat gene. / Richardson, Jeanne M.; Pessin, Jeffrey E.

In: Journal of Biological Chemistry, Vol. 268, No. 28, 05.10.1993, p. 21021-21027.

Research output: Contribution to journalArticle

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