Regulation of lipocalin-type prostaglandin D synthase gene expression by Hes-1 through E-box and interleukin-1 beta via two NF-kappa B elements in rat leptomeningeal cells

• Published in: The Journal of biological chemistry 2003-02, Vol.278 (8), p.6018-6026
• Main Authors: Fujimori, Ko, Fujitani, Yasushi, Kadoyama, Keiichi, Kumanogoh, Haruko, Ishikawa, Koichi, Urade, Yoshihiro
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Basic Helix-Loop-Helix Transcription Factors; Cerebral Cortex - enzymology; DNA Primers; Gene Expression Regulation, Enzymologic; Homeodomain Proteins - metabolism; Humans; Interleukin-1 - physiology; Intramolecular Oxidoreductases - genetics; Lipocalins; Meninges - enzymology; Molecular Sequence Data; NF-kappa B - metabolism; Rats; Repressor Proteins - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; Sequence Alignment; Sequence Homology, Nucleic Acid; Transcription Factor HES-1; Transcription, Genetic
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M208288200

The promoter function of the rat lipocalin-type prostaglandin D synthase (L-PGDS) gene was characterized in primary cultures of leptomeningeal cells prepared from the neonatal rat brain. Luciferase reporter assays with deletion and site-directed mutation of the promoter region (-1250 to +77) showed that an AP-2 element at -109 was required for activation and an E-box at +57, for repression. Binding of nuclear factors to each of these cis-elements was demonstrated by an electrophoretic mobility shift assay. Several components of the Notch-Hes signaling pathway, Jagged, Notch1, Notch3, and Hes-1, were expressed in the leptomeningeal cells. Human Hes-1 co-expressed in the leptomeningeal cells bound to the E-box of the rat L-PGDS gene, and repressed the promoter activity of the rat L-PGDS gene in a dose-dependent manner. The L-PGDS gene expression was up-regulated slowly by interleukin-1 beta to the maximum level at 24 h. The reporter assay with deletion and mutation revealed that two NF-kappa B elements at -1106 and -291 were essential for this up-regulation. Binding of two NF-kappa B subunits, p65 and c-Rel, to these two NF-kappa B elements occurred after the interleukin-1 beta treatment. Therefore, the L-PGDS gene is the first gene identified as the target for the Notch-Hes signal through the E-box among a variety of genes involved in the prostanoid biosynthesis, classified to the lipocalin family, and expressed in the leptomeninges. Moreover, the L-PGDS gene is a unique gene that is activated slowly by the NF-kappa B system.