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Papers In Press, published online ahead of print October 19, 2001
School of Life Sciences, University of Dundee, Dundee, Scotland DD1 5EH
Corresponding Author: v.poli{at}dundee.ac.uk
Prostaglandins are important mediators of activated macrophage functions and their inducible synthesis is mediated by Cycloxygenase-2 (COX-2). Here, we make use of the murine macrophage cells RAW264 as well as of immortalised macrophages derived from mice deficient for the transcription factor C/EBP
J. Biol. Chem, 10.1074/jbc.M108282200
Submitted on August 28, 2001
Revised on October 19, 2001
Accepted on October 18, 2001
The induction of cyclooxygenase-2 mRNA in macrophages is biphasic and requires both C/EBP
and C/EBP
transcription factors
to explore the molecular mechanisms regulating COX-2 induction in activated macrophages. We demonstrate that lipopolysaccharide-mediated COX-2 mRNA induction is biphasic. The initial phase is independent of de novo protein synthesis, correlates with CREB activation, is inhibited by treatments that abolish CREB phosphorylation and reduce NF-kB-mediated gene activation, and requires the presence of the transcription factor C/EBP. On the other hand, C/EBP
appears to be essential in addition to C/EBP to effect the second phase of COX-2 gene transcription, which is important for maintaining the induced state and requires de novo protein synthesis. Indeed, both phases of COX-2 induction were defective in C/EBP-/- macrophages. Moreover, the synthesis of C/EBP was increased dramatically by treatment with lipopolysaccharide and, like COX-2 induction, repressed by combined inhibition of the MAPK and of the SAPK2/p38 cascades. Taken together, these data identify CREB, NF-kB and both C/EBP and as key factors in co-ordinately orchestrating transcription from the COX-2 promoter in activated macrophages.
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