Activator protein-1 mediates shear stress-induced prostaglandin D synthase gene expression in vascular endothelial cells

Megumi Miyagi, Yoshikazu Miwa, Fumi Takahashi-Yanaga, Sachio Morimoto, Toshiyuki Sasaguri

Research output: Contribution to journalArticlepeer-review

33 Citations (Scopus)

Abstract

Objective - We attempted to determine the molecular mechanism of fluid shear stress-induced lipocalin-type prostaglandin D synthase (L-PGDS) expression in vascular endothelial cells. Methods and Results - We examined the promoter region of the L-PGDS gene by loading laminar shear stress (20 dyne/cm 2), using a parallel-plate flow chamber, on endothelial cells transfected with luciferase reporter vectors containing the 5′-flanking regions of the human L-PGDS gene. A deletion mutant analysis revealed that a shear stress-responsive element resided in the region between -2607 and -2523 bp. A mutation introduced into the putative binding site for activator protein-1 (AP-1) within this region eliminated the response to shear stress. In an electrophoretic mobility shift assay, shear stress stimulated nuclear protein binding to the AP-1 binding site, which was supershifted by antibodies to c-Fos and c-Jun. Shear stress elevated the c-Jun phosphorylation level in a time-dependent manner, similar to that of L-PGDS gene expression. SP600125, a c-Jun N-terminal kinase inhibitor, decreased the c-Jun phosphorylation, DNA binding of AP-1, and L-PGDS expression induced by shear stress. Additionally, an mRNA chase experiment using actinomycin D demonstrated that shear stress did not stabilize L-PGDS mRNA. Conclusions - Shear stress induces L-PGDS expression by transcriptional activation through the AP-1 binding site.

Original languageEnglish
Pages (from-to)970-975
Number of pages6
JournalArteriosclerosis, thrombosis, and vascular biology
Volume25
Issue number5
DOIs
Publication statusPublished - May 2005

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine

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