Transcription factor AP-2 functions as a repressor that contributes to the liver-specific expression of serum amyloid A1 gene.
We previously identified transcription factor AP-2 as the nuclear factor that interacts with the tissue-specific repressor element in the rat serum amyloid A1 (SAA1) promoter. In this report, we provide evidence for a second AP-2-binding site and show that both AP-2 sites participate in mediating the transcription repression of SAA1 promoter. This proximal AP-2 site overlaps with the NFkappaB-binding site known to be essential for SAA1 promoter activity. Protein binding competition experiments demonstrated that AP-2 and NFkappaB binding to these overlapping sites were mutually exclusive. Furthermore, the addition of AP-2 easily displaced prebound NFkappaB, whereas NFkappaB could not displace AP-2. These results thus suggest that one mechanism by which AP-2 negatively regulates SAA1 promoter activity may be by antagonizing the function of NFkappaB. Consistent with a repression function, transient expression of AP-2 in HepG2 cells inhibited conditioned medium-induced SAA1 promoter activation. This inhibition was dependent on functional AP-2-binding sites, since mutation of AP-2-binding sites abolished inhibitory effects of AP-2 in HepG2 cells as well as resulted in derepression of the SAA1 promoter in HeLa cells. In addition to SAA1, we found that several other liver gene promoters also contain putative AP-2-binding sites. Some of these sequences could specifically inhibit AP-2.DNA complex formation, and for the human complement C3 promoter, overexpression of AP-2 also could repress its cytokine-mediated activation. Finally, stable expression of AP-2 in hepatoma cells significantly reduced the expression of endogenous SAA, albumin, and alpha-fetoprotein genes. Taken together, our results suggest that AP-2 may function as a transcription repressor to inhibit the expression of not only SAA1 gene but also other liver genes in nonhepatic cells.
Department of Biochemistry and Molecular Biology, Program in Genes and Development, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
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D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000502:physiology*; D000685:Serum Amyloid A Protein / Q000502:physiology*; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000502:physiology*
D004268:DNA-Binding Proteins; D000685:Serum Amyloid A Protein; D050656:Transcription Factor AP-2; D014157:Transcription Factors