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Originally published In Press as doi:10.1074/jbc.M001430200 on June 1, 2000

J. Biol. Chem., Vol. 275, Issue 34, 26245-26251, August 25, 2000
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The Basic Helix-Loop-Helix Transcription Factors Myogenin and Id2 Mediate Specific Induction of Caveolin-3 Gene Expression during Embryonic Development*

Carola H. BiedererDagger §, Stefan J. RiesDagger §, Markus Moser||, Monica Florio**, Mark A. Israel**, Frank McCormickDagger Dagger Dagger , and Reinhard Buettner||§§

From the Dagger  University of California, San Francisco, School of Medicine, Cancer Research Institute, San Francisco, California 94143-0128, the || Institute of Pathology, University Hospital RWTH Aachen, D-52074 Aachen, Germany, and the ** University of California, San Francisco, Preuss Laboratory for Molecular Neuro-Oncology, Brain Tumor Research Center, Department of Neurological Surgery, San Francisco, California 94143-0520

Caveolin-3 protein is the only member of the caveolin family that shows a unique muscle-specific expression pattern, and loss of its functional activity causes muscular dystrophy. Caveolin-3 mRNA levels are dramatically increased during the formation of myotubes in the C2C12 cell line. In this study, we characterized the human caveolin-3 5'-flanking region. Promoter analyses demonstrate that the proximal E box element serves as a myogenin binding site and is both necessary and sufficient to control caveolin-3 gene transcription. Transient transfection assays indicated that overexpression of myogenin activates caveolin-3 reporter gene expression, whereas Id2 overexpression inhibited caveolin-3 promoter activation by myogenin. A mutant Id2 protein lacking the HLH domain was not capable of suppressing myogenin-mediated activation. Determination of caveolin-3 transcript distribution patterns in vivo revealed that mRNA was first detectable at day 10 of gestation in the developing somites and heart. Caveolin-3 protein in myoblasts and myotubes was expressed in both the plasma membrane and vesicular structures. During skeletal myogenesis the level of Id2, an inhibitor of differentiation, decreases, allowing the induced basic helix-loop-helix transcription factor myogenin to form transcriptionally active heterodimers that bind to the caveolin-3 promoter and thereby mediate its transcription.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF204690.

§ These authors contributed equally to this work.

Recipient of a Deutscher Akademischer Austausch Dieust (DAAD) fellowship as part of the Gemeinsames Hochschulsonderprogramm III von Bund und Laendern.

Dagger Dagger Supported by the David A. Wood Foundation.

§§ To whom correspondence should be addressed: Inst. of Pathology, University Hospital, RWTH Aachen, Pauwelsstrasse 30, D-52074 Aachen, Germany. Tel.: 49-241-8089280; Fax: 49-241-8888439; E-mail: Buettner@pat.rwth-aachen.de.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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