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J. Biol. Chem., Vol. 276, Issue 28, 26122-26131, July 13, 2001
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From the ¶ Department of Veterans Affairs,
Nashville, Tennessee 37212, The CXC chemokine, melanoma growth stimulatory
activity/growth-regulated protein, CXCL1 is an important modulator of
inflammation, wound healing, angiogenesis, and tumorigenesis.
Transcription of CXCL1 is regulated through several cis-acting elements
including Sp1, NF-
The Role of CDP in the Negative Regulation of
CXCL1 Gene Expression*
,,,¶
Vanderbilt University
School of Medicine, Department of Cancer Biology,
Nashville, Tennessee 37232, and the § Molecular Oncology
Group, McGill University, Montreal, Quebec H3A 1A1, Canada
B, and an element that lies immediately upstream
of the NF-B element, the immediate upstream region (IUR). A
transcription element data base search indicated that the IUR element
contains a binding site for the transcriptional repressor, human
CUT homeodomain protein/CCAAT displacement protein (CDP). It is
shown here that in electrophoretic mobility shift assays, complexes
obtained with the IUR oligonucleotide probe are supershifted by
anti-CDP antibodies and that a CDP polypeptide containing a high
affinity DNA binding domain binds to the sequence GGGATCGATC in the IUR
element. In Southwestern blot analyses, oligonucleotides containing the
wild-type IUR sequence, but not a mutant oligonucleotide with
substitutions in the GGGATCGATC sequence, bind a 170-180-kDa protein.
Furthermore, overexpression of the CDP protein blocks CXCL1
promoter activity in reporter gene assays, whereas overexpression of an
antisense CDP construct leads to a significant increase in
CXCL1 promoter activity. Mutations in the IUR element,
which map in the putative CDP-binding site, inhibit the binding of CDP
to the IUR element and favor increased transcription from the
CXCL1 promoter. Based on these results, we propose that
transcriptional regulation of the CXCL1 gene is mediated in
part by CDP, which could play an important role in inflammatory
processes and tumorigenesis.
*
This work was supported by NCI Grant CA 56704 from the
National Institutes of Health (to A. R.), a Senior Career Scientist award (to A. R.) from the Department of Veterans Affairs, an
Historically Black Colleges and Universities/Department of
Veterans Affairs grant, and NCI Grant CA 68485 from the National
Institutes of Health.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. Tel.:
615-343-7777; Fax: 615-343-4539; E-mail:
ann.richmond@mcmail.vanderbilt.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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