Cooperative activation of lipocalin-type prostaglandin D synthase gene expression by activator protein-2beta in proximal promoter and upstream stimulatory factor 1 within intron 4 in human brain-derived TE671 cells.
We investigated the activation mechanism of gene expression of lipocalin-type prostaglandin D synthase (L-PGDS) in human brain-derived TE671 cells. Reporter analyses of constructs carrying various lengths of the promoter region and intron 1 to 6, or 3'-untranslated region of the human L-PGDS gene demonstrated that one atypical E-box (aE-box) at +2569 in intron 4 was critical for transactivation of the gene. The aE-box inside the intron 4 functioned as an enhancer element in both directions and in a cell-type specific manner in TE671 cells. Yeast one-hybrid screening revealed that upstream stimulatory factor (USF) 1 bound to the aE-box. Expression of exogenous USF1 induced the endogenous L-PGDS expression in TE671 cells, whereas administration of USF1 siRNA suppressed L-PGDS expression. Binding of USF1 to the aE-box was confirmed by performing electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Furthermore, USF1-mediated transcriptional activation was dependent upon activator protein (AP)-2beta binding to the AP-2 element at position -98 in the proximal promoter region of human L-PGDS gene. These results indicate that L-PGDS gene expression in TE671 cells was activated by USF1 through the aE-box within intron 4 and cooperatively by AP-2beta in the promoter in a cell-type-specific manner.
Fujimori
Ko
K
Department of Molecular Behavioral Biology, Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka, Japan.
Urade
Yoshihiro
Y
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D066298:In Vitro Techniques; D019746:Intramolecular Oxidoreductases / Q000235:genetics*; D007438:Introns; D054834:Lipocalins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D050656:Transcription Factor AP-2 / Q000378:metabolism*; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques; D051720:Upstream Stimulatory Factors / Q000235:genetics / Q000378:metabolism*
D054834:Lipocalins; D034741:RNA, Small Interfering; D050656:Transcription Factor AP-2; C497181:USF1 protein, human; D051720:Upstream Stimulatory Factors; D004247:DNA; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase