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J. Biol. Chem., Vol. 279, Issue 44, 45423-45432, October 29, 2004

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The CCAAT Enhancer-binding Protein (C/EBP) and Nrf1 Interact to Regulate Dentin Sialophosphoprotein (DSPP) Gene Expression during Odontoblast Differentiation^*

Karthikeyan Narayanan, Amsaveni Ramachandran, Mathew Craig Peterson, Jianjun Hao, Anne-Brit Kolstø, Alan D. Friedman¶, and Anne George||

From the Department of Oral Biology (M/C 690), University of Illinois at Chicago, Chicago, Illinois 60612, the Biotechnology Centre of Oslo, University of Oslo, Oslo 0316, Norway, and the ^¶Department of Pediatric Oncology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21287

Terminal differentiation of odontoblasts, the principal cells in dentin formation, proceeds by synthesis of type I collagen and noncollagenous proteins. DSP and DPP are specific markers for terminally differentiated odontoblasts and are encoded by a single gene DSPP (dentin sialophosphoprotein). In an attempt to understand the molecular mechanisms required for tissue-specific expression of the DSPP gene, we have identified a novel interaction between two bZIP transcription factors, Nrf1 and the CCAAT enhancer-binding protein (C/EBP). This interaction was confirmed by both immunoprecipitation and chromatin immunoprecipitation assays. In undifferentiated odontoblasts, Nrf1 and C/EBP repress DSPP promoter activity individually and synergistically by cooperatively interacting with each other. This mutual interaction is facilitated by the bZIP domains in both the proteins. The repression domain in both Nrf1 and C/EBP was determined, and deletion of this domain abolished transcriptional repression. In fully differentiated odontoblasts, the loss of interaction between Nrf1 and C/EBP results in an increased DSPP transcription. Further, this interaction was found to be dependent on phosphorylation at Ser⁵⁹⁹ of Nrf1. Thus, the physical interaction between Nrf1 and C/EBP provide a novel mechanism for the transcriptional regulation of DSPP in odontoblasts.

Received for publication, May 6, 2004 , and in revised form, August 11, 2004.

^* This study was supported by National Institutes of Health Grants DE 13836 and DE 11657. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplementary figures.

^|| To whom correspondence should be addressed. Tel.: 312-413-0738; Fax: 312-996-6044; E-mail: anneg{at}uic.edu.

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