The Induction of Cyclooxygenase-2 mRNA in Macrophages Is Biphasic and Requires both CCAAT Enhancer-binding protein β (C/EBPβ) and C/EBPδ Transcription Factors

Matilde Caivano, Barbara Gorgoni, Philip Cohen, Valeria Poli

    Research output: Contribution to journalArticle

    132 Citations (Scopus)

    Abstract

    Prostaglandins are important mediators of activated macrophage functions, and their inducible synthesis is mediated by cyclooxygenase-2 (COX-2). Here, we make use of the murine macrophage cells RAW264 as well as of immortalized macrophages derived from mice deficient for the transcription factor CCAAT enhancer-binding protein β (C/EBPβ) 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 cAMP-response element-binding protein (CREB) activation, is inhibited by treatments that abolish CREB phosphorylation and reduce NF-κB-mediated gene activation, and requires the presence of the transcription factor C/EBPβ. On the other hand, C/EBPδ appears to be essential in addition to C/EBPβ 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/EBPβ-/- macrophages. Moreover, the synthesis of C/EBPδ 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-κB, and both C/EBPβ and -δ as key factors in coordinately orchestrating transcription from the COX-2 promoter in activated macrophages.

    Original languageEnglish
    Pages (from-to)48693-48701
    Number of pages9
    JournalJournal of Biological Chemistry
    Volume276
    Issue number52
    DOIs
    Publication statusPublished - 28 Dec 2001

    Fingerprint

    CCAAT-Enhancer-Binding Proteins
    Macrophages
    Cyclooxygenase 2
    Transcription Factors
    Messenger RNA
    Cyclic AMP Response Element-Binding Protein
    Transcription
    Lipopolysaccharides
    Mitogen-Activated Protein Kinase 11
    Genes
    Chemical activation
    Phosphorylation
    p38 Mitogen-Activated Protein Kinases
    Transcriptional Activation
    Prostaglandins
    Proteins

    Cite this

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    title = "The Induction of Cyclooxygenase-2 mRNA in Macrophages Is Biphasic and Requires both CCAAT Enhancer-binding protein β (C/EBPβ) and C/EBPδ Transcription Factors",
    abstract = "Prostaglandins are important mediators of activated macrophage functions, and their inducible synthesis is mediated by cyclooxygenase-2 (COX-2). Here, we make use of the murine macrophage cells RAW264 as well as of immortalized macrophages derived from mice deficient for the transcription factor CCAAT enhancer-binding protein β (C/EBPβ) 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 cAMP-response element-binding protein (CREB) activation, is inhibited by treatments that abolish CREB phosphorylation and reduce NF-κB-mediated gene activation, and requires the presence of the transcription factor C/EBPβ. On the other hand, C/EBPδ appears to be essential in addition to C/EBPβ 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/EBPβ-/- macrophages. Moreover, the synthesis of C/EBPδ 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-κB, and both C/EBPβ and -δ as key factors in coordinately orchestrating transcription from the COX-2 promoter in activated macrophages.",
    author = "Matilde Caivano and Barbara Gorgoni and Philip Cohen and Valeria Poli",
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    The Induction of Cyclooxygenase-2 mRNA in Macrophages Is Biphasic and Requires both CCAAT Enhancer-binding protein β (C/EBPβ) and C/EBPδ Transcription Factors. / Caivano, Matilde; Gorgoni, Barbara; Cohen, Philip; Poli, Valeria.

    In: Journal of Biological Chemistry, Vol. 276, No. 52, 28.12.2001, p. 48693-48701.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The Induction of Cyclooxygenase-2 mRNA in Macrophages Is Biphasic and Requires both CCAAT Enhancer-binding protein β (C/EBPβ) and C/EBPδ Transcription Factors

    AU - Caivano, Matilde

    AU - Gorgoni, Barbara

    AU - Cohen, Philip

    AU - Poli, Valeria

    PY - 2001/12/28

    Y1 - 2001/12/28

    N2 - Prostaglandins are important mediators of activated macrophage functions, and their inducible synthesis is mediated by cyclooxygenase-2 (COX-2). Here, we make use of the murine macrophage cells RAW264 as well as of immortalized macrophages derived from mice deficient for the transcription factor CCAAT enhancer-binding protein β (C/EBPβ) 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 cAMP-response element-binding protein (CREB) activation, is inhibited by treatments that abolish CREB phosphorylation and reduce NF-κB-mediated gene activation, and requires the presence of the transcription factor C/EBPβ. On the other hand, C/EBPδ appears to be essential in addition to C/EBPβ 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/EBPβ-/- macrophages. Moreover, the synthesis of C/EBPδ 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-κB, and both C/EBPβ and -δ as key factors in coordinately orchestrating transcription from the COX-2 promoter in activated macrophages.

    AB - Prostaglandins are important mediators of activated macrophage functions, and their inducible synthesis is mediated by cyclooxygenase-2 (COX-2). Here, we make use of the murine macrophage cells RAW264 as well as of immortalized macrophages derived from mice deficient for the transcription factor CCAAT enhancer-binding protein β (C/EBPβ) 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 cAMP-response element-binding protein (CREB) activation, is inhibited by treatments that abolish CREB phosphorylation and reduce NF-κB-mediated gene activation, and requires the presence of the transcription factor C/EBPβ. On the other hand, C/EBPδ appears to be essential in addition to C/EBPβ 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/EBPβ-/- macrophages. Moreover, the synthesis of C/EBPδ 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-κB, and both C/EBPβ and -δ as key factors in coordinately orchestrating transcription from the COX-2 promoter in activated macrophages.

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