Dioxin increases C/EBPbeta transcription by activating cAMP/protein kinase A.
The environmental pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD = dioxin) has been shown to increase the expression of C/EBPbeta. The modulated expression of C/EBPbeta has been suggested to be associated with toxic responses of TCDD such as wasting syndrome, diabetes, and inhibition of adipocyte differentiation. This study focused on the regulatory mechanism of TCDD-mediated transcriptional activation of C/EBPbeta. Elevated C/EBPbeta mRNA and protein levels in mouse embryonic fibroblasts (C3H10T(1/2)) and in mouse hepatoma cells (Hepa1c1c7) were correlated with increased binding affinity of the C/EBPbeta protein. Transfection studies with different deletion constructs of the CCAAT/enhancer-binding protein promoter indicated that a small region located 60-120 bp upstream of the start site of transcription is required for activation of the C/EBPbeta gene by TCDD in both cell lines tested. Further analysis using mutation constructs of the C/EBPbeta promoter demonstrated that activation of the C/EBPbeta promoter is mediated through incomplete cAMP-response element-binding protein (CREB) sites located close to the TATA box of the C/EBPbeta gene. The protein kinase A (PKA) inhibitor H89 completely blocks the TCDD-dependent effect on C/EBPbeta promoter activity, indicating that TCDD activates CREB binding via a cAMP/PKA pathway, which is supported by the increased cAMP level and PKA activity observed after TCDD treatment. Gel shift analyses demonstrated that CREB itself binds to the putative CREB motif that mediates the TCDD-dependent effect on C/EBPbeta gene transcription. Cotransfection experiments with CREB and PKA expression plasmids further supported our conclusions that the TCDD-dependent effect on C/EBPbeta transcription is mediated via PKA-dependent CREB activation.
Vogel
Christoph F A
CF
Department of Environmental Toxicology, University of California, Davis, California 95616, USA.
Sciullo
Eric
E
Park
Sujin
S
Liedtke
Christian
C
Trautwein
Christian
C
Matsumura
Fumio
F
D000818:Animals; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000096:biosynthesis* / Q000235:genetics; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D051379:Mice; D000072317:Polychlorinated Dibenzodioxins / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D015398:Signal Transduction / Q000187:drug effects; D013723:Teratogens / Q000494:pharmacology*; D015533:Transcriptional Activation / Q000187:drug effects*
D022782:CCAAT-Enhancer-Binding Protein-beta; D017362:Cyclic AMP Response Element-Binding Protein; D000072317:Polychlorinated Dibenzodioxins; D013723:Teratogens; D017868:Cyclic AMP-Dependent Protein Kinases