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

Gene. 2007 Aug 1;397(1-2):143-52. doi: 10.1016/j.gene.2007.04.029. Epub 2007 May 3.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites / genetics
  • Brain / metabolism
  • Cell Line
  • DNA / genetics
  • DNA / metabolism
  • Gene Expression Regulation, Enzymologic
  • Humans
  • In Vitro Techniques
  • Intramolecular Oxidoreductases / genetics*
  • Introns
  • Lipocalins
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • RNA, Small Interfering / genetics
  • Sequence Homology, Nucleic Acid
  • Transcription Factor AP-2 / metabolism*
  • Transcriptional Activation
  • Two-Hybrid System Techniques
  • Upstream Stimulatory Factors / genetics
  • Upstream Stimulatory Factors / metabolism*

Substances

  • Lipocalins
  • RNA, Small Interfering
  • Transcription Factor AP-2
  • USF1 protein, human
  • Upstream Stimulatory Factors
  • DNA
  • Intramolecular Oxidoreductases
  • prostaglandin R2 D-isomerase