The CXC chemokine, melanoma growth stimulatory activity/growth-regulated pr
otein, CXCL1 is an important modulator of inflammation, wound healing, angi
ogenesis, and tumorigenesis, Transcription of CXCL1 is regulated through se
veral cia-acting elements including Spl, NF-kappaB, and an element that lie
s immediately upstream of the NF-kappaB element, the immediate upstream reg
ion (IUR), A transcription element data base search indicated that the IUR
element contains a binding site for the transcriptional repressor, human CU
T homeodomain protein/CCAAT displacement protein (CDP), It is shown here th
at in electrophoretic mobility shift assays, complexes obtained with the IU
R 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, olig
onucleotides containing the wild-type IUR sequence, but not a mutant oligon
ucleotide with substitutions in the GGGATCGATC sequence, bind a 170-180-kDa
protein. Furthermore, overexpression of the CDP protein blocks CXCL1 promo
ter activity in reporter gene assays, whereas overexpression of an antisens
e CDP construct leads to a significant increase in CXCL1 promoter activity.
Mutations in the IUR element, which map in the putative CUP-binding site,
inhibit the binding of CDP to the IUR element and favor increased transcrip
tion from the CXCL1 promoter. Based on these results, we propose that trans
criptional regulation of the CXCL1 gene is mediated in part by CDP, which c
ould play an important role in inflammatory processes and tumorigenesis.