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J. Biol. Chem., Vol. 277, Issue 35, 31526-31533, August 30, 2002
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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 1
CCAAT/Enhancer-binding Proteins
and 
Mediate the
Repression of Gene Transcription of Cartilage-derived Retinoic
Acid-sensitive Protein Induced by Interleukin-1*
(IL-1). To investigate
the transcriptional mechanism, by which CD-RAP expression is suppressed
by IL-1, deletion constructs of the mouse CD-RAP promoter were
transfected into rat chondrocytes treated with or without IL-1. The
results revealed an IL-1-responsive element located between 
2138
and 2068 bp. As this element contains a CAAT/enhancer-binding protein (C/EBP) motif, the function of C/EBP and C/EBP
was examined. IL-1 stimulated the expression of C/EBP and -, and the direct binding of C/EBP 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-1. 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/EBP and -
may play an important role in the IL-1-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.
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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