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(Received for publication, July 30, 1996, and in revised form, March 3, 1997)
From the Interferon-
Volume 272, Number 23,
Issue of June 6, 1997
pp. 14899-14907
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
and Tumor Necrosis Factor-
in
Transcriptional Activation Is Mediated by Cooperation between Signal
Transducer and Activator of Transcription 1 and Nuclear Factor
B
,
Department of Immunology, The Cleveland
Clinic Foundation, Cleveland, Ohio 44195 and the ¶ Center for
Immunology, Department of Pathology, Washington University School of
Medicine, St. Louis, Missouri 63110
(IFN) and tumor necrosis
factor-
(TNF) cooperate to induce the expression of many gene
products during inflammation. The present report demonstrates that a
portion of this cooperativity is mediated by synergism between two
distinct transcription factors: signal transducer and activator of
transcription 1 (STAT1) and nuclear factor 
B (NF-B). IFN and
TNF synergistically induce expression of mRNAs encoding
interferon regulatory factor-1 (IRF-1), intercellular adhesion
molecule-1, Mig (monokine induced by -interferon), and RANTES
(regulated on activation normal
T cell expressed and secreted) in normal but not STAT1-deficient mouse
fibroblasts, indicating a requirement for STAT1. Transient transfection
assays in fibroblasts using site-directed mutants of a 1.3-kilobase
pair sequence of the IRF-1 gene promoter revealed that the synergy was
dependent upon two sequence elements; a STAT binding element and a B
motif. Artificial constructs containing a single copy of both a STAT
binding element and a B motif linked to the herpes virus thymidine
kinase promoter were able to mediate synergistic response to IFN and
TNF; such response varied with both the relative spacing and the
specific sequence of the regions between these two sites. Cooperatively
responsive sequence constructs bound both STAT1 and NF-B in
nuclear extracts prepared from IFN- and/or TNF-stimulated
fibroblasts, although binding of individual factors was not
cooperative. Thus, the frequently observed synergy between IFN and
TNF in promoting inflammatory response depends in part upon
cooperation between STAT1 and NF-B, which is most likely mediated
by their independent interaction with one or more components of the
basal transcription complex.
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