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-Stimulated Monocyte Chemoattractant Protein-1 Gene Transcription1
*
Department of Medicine, University of Texas Health Science Center, San Antonio, TX 78284;
Division of Oral Biology, Boston University School of Dental Medicine, Boston, MA 02118; and
Department of Medicine, University of Hamburg, Hamburg, Germany
Monocyte chemoattractant protein-1 (MCP-1) is induced in chronic
osseous inflammation, and is temporally and spatially correlated with
monocyte recruitment. We investigated the mechanism of MCP-1 regulation
in a human osteoblastic cell line in response to IFN-
, a potent
mediator of the immune inflammatory response. Nuclear run-on and
stability studies demonstrated that IFN- stimulated MCP-1
transcription and did not enhance mRNA stabilization. Using MCP-1
promoter/reporter gene constructs, we determined that IFN--enhanced
MCP-1 transcription is regulated by a 29-bp element located at -227
relative to the ATG start codon. This element contains a 13-bp CT-rich
sequence (GCTTCCCTTTCCT) adjacent to a IFN- activation site (GAS).
Since deletion of the CT sequence enhanced both the magnitude and
duration of IFN--stimulated, GAS-mediated transcription, we have
termed it the IFN response-inhibitory sequence (IRIS). The combined
IRIS/GAS sequence is highly conserved in mouse, rat, and bovine MCP-1
genes. In gel-shift assays, nuclear extracts from IFN--stimulated
osteoblastic cells formed two specific inducible bands with labeled
IRIS/GAS DNA. Both bands were supershifted by anti-STAT1 Abs, but
not by Abs to STAT2, p48(ISGF-3), IFN-regulatory factor-1, or
IFN-regulatory factor-2. Formation of one of the bands required the
presence of the IRIS moiety. IRIS/GAS DNA also formed a number of
specific complexes with constitutively expressed factors, none of which
were affected by the above Abs. These studies establish a mechanism for
IFN--stimulated MCP-1 expression and identify a complex element that
regulates MCP-1 gene transcription.
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