Protein kinase C activates human lipocalin-type prostaglandin D synthase gene expression through de-repression of notch-HES signaling and enhancement of AP-2 beta function in brain-derived TE671 cells

J Biol Chem. 2005 May 6;280(18):18452-61. doi: 10.1074/jbc.M411755200. Epub 2005 Mar 2.


Here we investigated the regulatory mechanism of lipocalin-type prostaglandin D synthase (L-PGDS) gene expression in human TE671 (medulloblastoma of cerebellum) cells. Reporter analysis of the promoter region from -730 to +75 of the human L-PGDS gene demonstrated that deletion or mutation of the N-box at -337 increased the promoter activity 220-300%. The N-box was bound by Hes-1, a mammalian homologue of Drosophila Hairy and enhancer of split, as examined by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Functional expression of the Notch intracellular domain significantly increased Hes-1 expression and decreased L-PGDS expression level in TE671 cells. Moreover, knock-down of Hes-1 mRNA by RNA interference significantly enhanced the L-PGDS mRNA level, indicating that the L-PGDS gene expression is repressed by the Notch-Hes signaling. When the AP-2 element at -98 of the promoter region was deleted or mutated, the promoter activity was drastically decreased to approximately 10% of normal. The AP-2 element was bound by AP-2beta dominantly expressed in TE671 cells, according to the results of electrophoretic mobility shift assay and chromatin immunoprecipitation assay. L-PGDS expression was induced by 12-O-tetradecanoylphorbol-13-acetate in TE671 cells, and this induction was inhibited by a protein kinase C inhibitor. Stimulation of TE671 cells with 12-O-tetradecanoylphorbol-13-acetate or transfection with protein kinase Calpha expression vector induced phosphorylation of Hes-1, inhibition of DNA binding of Hes-1 to the N-box, and activation of the AP-2beta function to up-regulate L-PGDS gene expression. These results reveal a novel transcriptional regulatory mechanism responsible for the high level expression of the human L-PGDS gene in TE671 cells.

Publication types

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

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors
  • Brain / metabolism
  • Brain / pathology
  • Cell Line, Tumor
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Enzyme Repression / drug effects
  • Enzyme Repression / genetics
  • Gene Expression Regulation, Enzymologic / physiology*
  • Homeodomain Proteins / antagonists & inhibitors
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Humans
  • Intramolecular Oxidoreductases / antagonists & inhibitors
  • Intramolecular Oxidoreductases / genetics*
  • Intramolecular Oxidoreductases / metabolism
  • Lipocalins
  • Medulloblastoma / enzymology*
  • Medulloblastoma / genetics
  • Medulloblastoma / pathology
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Protein Kinase C / physiology*
  • Protein Structure, Tertiary / genetics
  • RNA Interference
  • RNA, Messenger / biosynthesis
  • Receptors, Notch
  • Repressor Proteins / antagonists & inhibitors
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / genetics*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transcription Factor AP-2
  • Transcription Factor HES-1
  • Transcription Factors / genetics
  • Transcription Factors / physiology*


  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • Lipocalins
  • Membrane Proteins
  • RNA, Messenger
  • Receptors, Notch
  • Repressor Proteins
  • TFAP2B protein, human
  • Transcription Factor AP-2
  • Transcription Factor HES-1
  • Transcription Factors
  • HES1 protein, human
  • Protein Kinase C
  • Intramolecular Oxidoreductases
  • prostaglandin R2 D-isomerase
  • Tetradecanoylphorbol Acetate