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Gene1
Pulmonary Center and Department of Pathology, Boston University School of Medicine, Boston MA 02118
Both lymphoid and myeloid cells express two related members of the
IFN regulatory factor (IRF) family of transcription factors,
specifically IRF-4 and IFN consensus binding protein (ICSBP or IRF-8).
We previously reported that macrophages express IRF-4 and in
combination with the ETS-like protein PU.1 can synergistically
activate a human IL-1
reporter gene. Here we report that this
synergy is mediated by a composite PU.1/IRF element located within an
upstream enhancer known to confer cytokine- and LPS-inducible
expression. In macrophages, synergistic activation of IL-1 reporter
gene expression was preferentially mediated by IRF-4, whereas IRF-4 and
ICSBP were equally capable of synergizing with PU.1 when coexpressed in
fibroblasts. Furthermore, coexpression of IRF-1 and IRF-2 dramatically
increased the capacity of both PU.1/IRF-4 and PU.1/ICSBP to induce
IL-1 reporter gene expression in fibroblasts. The additional synergy
observed with IRF-1 and IRF-2 coexpression is mediated by a region of
DNA distinct from either the IL-1 enhancer or promoter. We also
assessed the capacity of these transcription factors to activate
endogenous IL-1 gene when overexpressed in human embryonic kidney
293 cells. Although ectopic expression of PU.1 alone was sufficient to
activate modest levels of IL-1 transcripts, endogenous IL-1
expression was markedly increased following coexpression of additional
IRF proteins. Thus, maximal expression of both a human IL-1 reporter
gene and the endogenous IL-1 gene was observed in cells that
coexpressed PU.1, IRF-4 (or ICSBP), IRF1, and IRF2. Together, our
observations suggest that these factors may function together as an
enhanceosome.
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