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Originally published In Press as doi:10.1074/jbc.M005929200 on August 18, 2000

J. Biol. Chem., Vol. 275, Issue 45, 34963-34967, November 10, 2000
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The Minimal Repression Domain of MBD2b Overlaps with the Methyl-CpG-binding Domain and Binds Directly to Sin3A*

Joern Boeke, Ole Ammerpohl, Simone Kegel, Udo Moehren, and Rainer RenkawitzDagger

From the Genetisches Institut der Justus-Liebig-Universität Gießen, Heinrich-Buff-Ring 58, D-35392 Gießen, Germany

Different mechanisms mediating methylation-dependent repression have been demonstrated. Two of these mechanisms play a role in the context of the granulocyte/macrophage-specific lysozyme gene: direct interference with DNA binding of the transcription factor GA-binding protein and deacetylation of histones. Besides enhancement in the unmethylated state, and transcriptional repression upon DNA methylation, the lysozyme downstream enhancer confers tissue-specific demethylation. Because both demethylation activity and repression ability have been attributed to the methyl-CpG-binding domain-containing protein MBD2, we analyzed this protein. The short form MBD2b binds to the methylated lysozyme enhancer and mediates transcriptional repression. MBD2b is capable of binding to the transcriptional repressor Sin3A. The interaction domain of Sin3A required for binding to MBD2b contains the paired amphipathic helix 3. We identified a minimal functional domain that confers both transcriptional repression as well as the interaction to Sin3A. In contrast to the functionally related proteins MeCP2 and MBD1, the repression domain of MBD2b overlaps with the methyl-CpG-binding domain.

* This work was supported by Deutsche Forschungsgemeinschaft Grant SFB397 and by the Fond der Chemischen Industrie.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.

Dagger To whom correspondence should be addressed. Tel.: 49-641-99-35460; Fax: 49-641-99-35469; E-mail: Rainer.Renkawitz@gen.bio. uni-giessen.de.

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