Cell Growth & Differentiation, Vol 8, Issue 1 23-34, Copyright © 1997 by American Association of Cancer Research

ARTICLES

Absence of MEF2 binding to the A/T-rich element in the muscle creatine kinase (MCK) enhancer correlates with lack of early expression of the MCK gene in embryonic mammalian muscle

S Ferrari, S Molinari, R Melchionna, MG Cusella-De Angelis, R Battini, L De Angelis, R Kelly and G Cossu
Dipartimento di Scienze Biomediche, Universita di Modena, Italy. sferrari@c220.unimo.it

During skeletal muscle development, different types of muscle fibers are generated, which express different combinations of muscle-specific gene products. For example, the muscle creatine kinase gene (MCK) is highly expressed in fetal but not embryonic myotubes. We performed transient transfections of CAT reporter constructs, driven by the MCK promoter with variable lengths of 5'-flanking sequence, into primary cultures of embryonic and fetal muscle cells. Reporter activity was observed in fetal but not embryonic muscle cells. We assayed the ability of nuclear extracts prepared from embryonic and fetal muscle and C2C12 myotubes to bind specific regulatory elements in the MCK enhancer. The profile of DNA/protein complexes resulting from electrophoretic mobility shift assays was qualitatively the same with all extracts used when the oligonucleotide probes represented the MCK-E-box, MHox site, CArG-box, and AP2 site. In contrast, no binding activity to the MEF2 site was observed with embryonic nuclear extract. Interestingly, MEF2 mRNAs and proteins were detected in both fetal and embryonic muscle, with the exception of the MEF2D1b isoform, which is restricted to fetal muscle. Furthermore, we found that protein phosphatase inhibitors included in the preparation of embryonic nuclear extracts or added to the medium of transfected embryonic myotubes can restore MEF2 DNA binding activity, as well as reporter activity driven by the MCK promoter and partial transcriptional activation of the endogenous MCK gene. We propose that phosphorylation of MEF2 regulates its activity and represents an important aspect of the mechanism controlling stage-specific transcription during skeletal myogenesis.

This article has been cited by other articles:

				C. Angelelli, A. Magli, D. Ferrari, M. Ganassi, V. Matafora, F. Parise, G. Razzini, A. Bachi, S. Ferrari, and S. Molinari Differentiation-dependent lysine 4 acetylation enhances MEF2C binding to DNA in skeletal muscle cells Nucleic Acids Res., February 11, 2008; 36(3): 915 - 928. [Abstract] [Full Text] [PDF]

				R. O. Balza Jr. and R. P. Misra Role of the Serum Response Factor in Regulating Contractile Apparatus Gene Expression and Sarcomeric Integrity in Cardiomyocytes J. Biol. Chem., March 10, 2006; 281(10): 6498 - 6510. [Abstract] [Full Text] [PDF]

				B. M. Scicchitano, L. Spath, A. Musaro, M. Molinaro, S. Adamo, and C. Nervi AVP Induces Myogenesis through the Transcriptional Activation of the Myocyte Enhancer Factor 2 Mol. Endocrinol., June 1, 2002; 16(6): 1407 - 1416. [Abstract] [Full Text] [PDF]

				M. G. Minasi, M. Riminucci, L. De Angelis, U. Borello, B. Berarducci, A. Innocenzi, A. Caprioli, D. Sirabella, M. Baiocchi, R. De Maria, et al. The meso-angioblast: a multipotent, self-renewing cell that originates from the dorsal aorta and differentiates into most mesodermal tissues Development, January 6, 2002; 129(11): 2773 - 2783. [Abstract] [Full Text] [PDF]

				G. Condorelli, U. Borello, L. De Angelis, M. Latronico, D. Sirabella, M. Coletta, R. Galli, G. Balconi, A. Follenzi, G. Frati, et al. Cardiomyocytes induce endothelial cells to trans-differentiate into cardiac muscle: Implications for myocardium regeneration PNAS, September 4, 2001; (2001) 191217898. [Abstract] [Full Text] [PDF]

				L. De Angelis, L. Berghella, M. Coletta, L. Lattanzi, M. Zanchi, M. Gabriella , Cusella-, C. Ponzetto, and G. Cossu Skeletal Myogenic Progenitors Originating from Embryonic Dorsal Aorta Coexpress Endothelial and Myogenic Markers and Contribute to Postnatal Muscle Growth and Regeneration J. Cell Biol., November 15, 1999; 147(4): 869 - 878. [Abstract] [Full Text] [PDF]

				D. R. Vyas, J. J. McCarthy, and R. W. Tsika Nuclear Protein Binding at the beta -Myosin Heavy Chain A/T-rich Element Is Enriched following Increased Skeletal Muscle Activity J. Biol. Chem., October 22, 1999; 274(43): 30832 - 30842. [Abstract] [Full Text] [PDF]

				A. Musaro and N. Rosenthal Maturation of the Myogenic Program Is Induced by Postmitotic Expression of Insulin-Like Growth Factor I Mol. Cell. Biol., April 1, 1999; 19(4): 3115 - 3124. [Abstract] [Full Text] [PDF]

				R. Passantino, V. Antona, G. Barbieri, P. Rubino, R. Melchionna, G. Cossu, S. Feo, and A. Giallongo Negative Regulation of beta  Enolase Gene Transcription in Embryonic Muscle Is Dependent upon a Zinc Finger Factor That Binds to the G-rich Box within the Muscle-specific Enhancer J. Biol. Chem., January 2, 1998; 273(1): 484 - 494. [Abstract] [Full Text] [PDF]

				G. Condorelli, U. Borello, L. De Angelis, M. Latronico, D. Sirabella, M. Coletta, R. Galli, G. Balconi, A. Follenzi, G. Frati, et al. From the Cover: Cardiomyocytes induce endothelial cells to trans-differentiate into cardiac muscle: Implications for myocardium regeneration PNAS, September 11, 2001; 98(19): 10733 - 10738. [Abstract] [Full Text] [PDF]