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Originally published In Press as doi:10.1074/jbc.M202815200 on June 18, 2002

J. Biol. Chem., Vol. 277, Issue 35, 31526-31533, August 30, 2002
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CCAAT/Enhancer-binding Proteins beta  and delta  Mediate the Repression of Gene Transcription of Cartilage-derived Retinoic Acid-sensitive Protein Induced by Interleukin-1beta *

Ken Okazaki, Jian Li, Hua Yu, Naoshi Fukui, and Linda J. SandellDagger

From the Department of Orthopaedic Surgery and Department of Cell Biology and Physiology, Washington University School of Medicine at Barnes-Jewish Hospital, St. Louis, Missouri 63110

Cartilage-derived retinoic acid-sensitive protein (CD-RAP) is a secreted protein expressed by chondrocytes; the expression is repressed by interleukin 1beta (IL-1beta ). To investigate the transcriptional mechanism, by which CD-RAP expression is suppressed by IL-1beta , deletion constructs of the mouse CD-RAP promoter were transfected into rat chondrocytes treated with or without IL-1beta . The results revealed an IL-1beta -responsive element located between -2138 and -2068 bp. As this element contains a CAAT/enhancer-binding protein (C/EBP) motif, the function of C/EBPbeta and C/EBPdelta was examined. IL-1beta stimulated the expression of C/EBPbeta and -delta , and the direct binding of C/EBPbeta to the C/EBP motif was confirmed. The -2251-bp CD-RAP promoter activity was down-regulated by co-transfection with C/EBP expression vectors. Mutation of the C/EBP motif abolished the inhibitory response to IL-1beta . Additionally, C/EBP expression vectors were found to down-regulate the construct containing the promoter and enhancer of the type II collagen gene. Finally, the enhancer factor, Sox9, was shown to bind adjacent to the C/EBP site competing with C/EBP binding. Taken together, these results suggest that C/EBPbeta and -delta may play an important role in the IL-1beta -induced repression of cartilage-specific proteins and that expression of matrix proteins will be influenced by the availability of positive and negative trans-acting factors.

* This work was supported by National Institutes of Health Grants R01AR45550 and R01AR36994.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: Dept. of Orthopaedic Surgery, Washington University School of Medicine at Barnes-Jewish Hospital, Mail Stop 90-34-674, 216 S. Kingshighway, St. Louis, MO 63110. Tel.: 314-454-7800; Fax: 314-454-5900; E-mail: sandelll@msnotes.wustl.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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