Enhancer Binding Protein Transcription Factor Research Articles

C/EBPβ controls mucosal fungal immunity through regulation of β-defensins (IRM11P.630)

Abstract Th17 cells and IL-17 play essential host-defensive roles in immunity to pathogens, particularly the commensal microbe Candida albicans. Although best known for activation of NF-κB, IL-17 potently activates CCAAT enhancer binding protein transcription factors, which are required for induction of many IL-17 target genes. C/EBPβ is translated into 3 distinct protein isoforms whose expression is regulated by the differential use of different translational initiation start sites, known as LAP* (38kDa), LAP (35kDa) and LIP (22kDa). We have shown that IL-17 induces the alternative translation of C/EBPβ, inducing a marked increase in LAP* and LIP. However, the functional significance of differential C/EBPβ isoform expression remains enigmatic. To determine the biological significance of C/EBPβ alternative translation, we used C/EBPβ-/- mice and a knockin that cannot generate the LAP isoform (C/EBPβM20A). We then assessed susceptibility of these mice to oral C. albicans infection. We found that C/EBPβ-/- but not C/EBPβM20 mice were highly susceptible to disease indicating that LAP is dispensable but C/EBPβ is required. In evaluating a panel of downstream genes to elucidate mechanisms of C/EBPβ-dependent immunity, the gene encoding β-defensin 3 (BD3) was the only IL-17 target gene strongly downregulated in susceptible C/EBPβ-/- mice. These results highlight a key role for C/EBP in antifungal immunity mediated by IL-17 and a previously unrecognized role for C/EBPβ in BD3 expression.

Opposing action of curcumin and green tea polyphenol in human keratinocytes

Persistent environmental insult can convert a normal cell into a cancer cell. However, various natural chemopreventive agents called antioxidants can retard this progression. We have recently explored the effects of several chemopreventive agents, including green tea polyphenol and curcumin, on normal human keratinocyte function. Our findings suggest that a bioactive polyphenol from green tea, (-)-epigallocatechin-3-gallate (EGCG), acts to increase involucrin gene expression, suggesting that EGCG treatment enhances normal human keratinocyte differentiation. Mechanistic studies indicate that EGCG alters mitogen-activated protein kinase cascade function to activate involucrin gene transcription via a Ras, MEKK1, MEK3, ERK1/2-p38delta cascade that targets AP1 and CAATT enhancer binding protein transcription factors. These findings suggest that EGCG may inhibit disease progression by promoting keratinocyte differentiation. Parallel studies indicate that not all antioxidants produce a similar response. Curcumin, an antioxidant derived from the turmeric, antagonizes the EGCG-dependent response by interfering in this signaling pathway. These studies suggest that different antioxidant may produce antagonistic effects in tissues.

Involvement of CCAAT enhancer binding protein transcription factors in the regulation of prostaglandin G/H synthase 2 expression by interleukin-1 in osteoblastic MC3T3-E1 cells.

Interleukin-1 (IL-1) stimulates prostaglandin production in bone by a rapid and transient activation of prostaglandin G/H synthase 2 (PGHS-2) gene expression. In osteoblastic MC3T3-E1 cells, IL-1 caused a transient increase in PGHS-2 messenger RNA (mRNA), which peaked at 2 h. IL-1 caused a 2- to 4-fold activation of a 371-base pair (bp) murine PGHS-2 promoter/luciferase construct in stable transfectants. This response mapped to a proximal promoter element(s) located between -150 and -40 bp. This region contains a putative CCAAT enhancer binding protein (C/EBP) site (centered at -135 bp), which shows enhanced binding of C/EBPbeta and C/EBPdelta by mobility shift analysis after IL-1 treatment. A transient cotransfection approach was used to examine the effects of C/EBPbeta and C/EBPdelta overexpression. IL-1 caused a maximal 3- to 7-fold stimulation of PGHS-2 promoter activity after 2.5 h. Overexpression of murine C/EBPbeta and C/EBPdelta caused a dose-dependent increase in basal and IL-1-stimulated luciferase activity. C/EBPdelta caused a greater enhancement of basal and IL-1-stimulated promoter activity than C/EBPbeta, suggesting that C/EBPdelta is a stronger transactivator. Overexpression of p20C/EBPbeta, a dominant negative inhibitor of C/EBP function, blocked the stimulation of PGHS-2 promoter activity by IL-1 and blocked the ability of overexpressed C/EBPbeta and C/EBPdelta to increase basal and IL-1-stimulated promoter activity. Mutagenesis of the C/EBP site reduced, but did not abolish, the stimulation of PGHS-2 promoter activity by IL-1 and blunted the effect of overexpressed C/EBPdelta on basal and IL-1-stimulated promoter activity. These results suggest an essential role for C/EBPbeta and C/EBPdelta in the induction of PGHS-2 gene expression by IL-1 in osteoblastic cells.