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A more recent version of this article appeared on May 1, 2003
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Submitted on July 31, 2002
Revised on December 20, 2002
Accepted on January 7, 2003
1 Cell Biology Unit, Institut de Génétique Humaine, 141, Route de la Cardonille, 34396 Montpellier cedex 05, France
2 Ecole Normale Supérieure de Lyon, Laboratoire de Biologie Moléculaire et Cellulaire, 46, allée d'Italie, 69364 Lyon cedex 07, France
3 Endothelial and Epithelial Cell Biology, Division of Cell Biology, Institute of Opthalmology, University College London Bath Street, London ECIV 9EL, England
* Corresponding author. E-mail address: af{at}acrux.igh.cnrs.fr.
We show here that the Distal Regulatory Region (DRR) of the mouse and human MyoD gene contains a conserved SRF binding CArG-like element. In electrophoretic mobility shift assays with myoblast nuclear extracts, this CArG sequence, although slightly divergent, bound two complexes containing respectively the transcription factor YY1 and SRF associated with the acetyltransferase CBP and members of C/EBP family. A single nucleotide mutation in the MyoD-CArG element suppressed binding of both SRF and YY1 complexes and abolished DRR enhancer activity in stably transfected myoblasts. This MyoD-CArG sequence is active in modulating endogeneous MyoD gene expression since microinjection of oligonucleotides corresponding to the MyoD-CArG sequence specifically and rapidly suppressed MyoD expression in myoblasts. In vivo, the expression of a transgenic construct comprising a minimal MyoD promoter fused to the DRR and 
-galactosidase was induced with the same kinetics as MyoD during mouse muscle regeneration. In contrast induction of this reporter was no longer seen in regenerating muscle from transgenic mice carrying a mutated DRR-CArG. These results show that an SRF binding CArG element present in MyoD gene DRR is involved in the control of MyoD gene expression in skeletal myoblasts and in mature muscle satellite cell activation during muscle regeneration.
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