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Molecular and Cellular Biology, April 2005, p. 3261-3275, Vol. 25, No. 8
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.8.3261-3275.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Myocyte Enhancer Factor 2 Activates Promoter Sequences of the Human AßH-J-J Locus, Encoding Aspartyl-ß-Hydroxylase, Junctin, and Junctate

Giordana Feriotto,¹ Alessia Finotti,² Pompeo Volpe,³ Susan Treves,⁴ Stefano Ferrari,⁵ Cecilia Angelelli,⁵ Francesco Zorzato,⁶ and Roberto Gambari^2*

Biotechnology Center,¹ Department of Biochemistry and Molecular Biology, Section of Molecular Biology,² Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara,⁶ Department of Biomedical Experimental Sciences, University of Padova, Padova,³ Department of Biomedical Sciences, University of Modena and Reggio Emilia, Modena, Italy,⁵ Departments of Anaesthesia and Research, Kantonsspital, Basel, Switzerland⁴

Received 23 August 2004/ Returned for modification 27 September 2004/ Accepted 30 December 2004

Alternative splicing of the locus AßH-J-J generates three functionally distinct proteins: an enzyme, AßH (aspartyl-ß-hydroxylase), a structural protein of the sarcoplasmic reticulum membrane (junctin), and an integral membrane calcium binding protein (junctate). Junctin and junctate are two important proteins involved in calcium regulation in eukaryotic cells. To understand the regulation of these two proteins, we identified and functionally characterized one of the two promoter sequences of the AßH-J-J locus. We demonstrate that the P2 promoter of the AßH-J-J locus contains (i) a minimal sequence localized within a region –159 bp from the transcription initiation site, which is sufficient to activate transcription of both mRNAs; (ii) sequences which bind known transcriptional factors such as those belonging to the myocyte enhancer factor 2 (MEF-2), MEF-3, and NF-B protein families; and (iii) sequences bound by unknown proteins. The functional characterization of the minimal promoter in C2C12 cells and in the rat soleus muscle in vivo model indicates the existence of cis elements having positive and negative effects on transcription. In addition, our data demonstrate that in striated muscle cells the calcium-dependent transcription factor MEF-2 is crucial for the transcription activity directed by the P2 promoter. The transcription directed by the AßH-J-J P2 promoter is induced by high expression of MEF-2, further stimulated by calcineurin and Ca2⁺/calmodulin-dependent protein kinase I, and inhibited by histone deacetylase 4.

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* Corresponding author. Mailing address: Dept. of Biochemistry and Molecular Biology, University of Ferrara, Via Borsari 46, 44100 Ferrara, Italy. Phone: 39 532 424443. Fax: 39 532 202723. E-mail: gam{at}unife.it.

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Molecular and Cellular Biology, April 2005, p. 3261-3275, Vol. 25, No. 8
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.8.3261-3275.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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