Transcriptional Regulation of Human Prostaglandin-endoperoxide Synthase-2 Gene by Lipopolysaccharide and Phorbol Ester in Vascular Endothelial Cells
INVOLVEMENT OF BOTH NUCLEAR FACTOR FOR INTERLEUKIN-6 EXPRESSION SITE AND cAMP RESPONSE ELEMENT (*)
- From the Department of Pharmacology, National Cardiovascular Center Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka 565, Japan
- § To whom correspondence should be addressed. Tel.: 81-6-833-5012 (ext. 2514); Fax: 81-6-872-7485.
Abstract
There exist two distinct isozymes of prostaglandin-endoperoxide synthase (PES). PES-2 mRNA is synergistically induced by lipopolysaccharide
(LPS) and 12-O-tetradecanoylphorbol-13-acetate (TPA) in bovine arterial endothelial cells. On the other hand, PES-1 mRNA is constitutively
expressed under these conditions. Therefore, the promoter activities of the human genes for PES-1 and −2 in bovine arterial
endothelial cells were examined. The 5′-flanking region of the human PES-2 gene (nucleotides −327 to +59) showed promoter
activity inducible by LPS and TPA using transient transfection analysis, whereas that of the PES-1 gene (nucleotides −1010
to +69) showed constitutive promoter activity. Destruction of both consensus sequences for the nuclear factor responsible
for the interleukin-6 expression (NF-IL6) site (nucleotides −132 to −124) and the cyclic AMP response element (CRE) (nucleotides
−59 to −53) of the human PES-2 gene markedly reduced the promoter activity (25%) of the PES-2 gene after combined treatment
with LPS and TPA, although single destruction of the NF-IL6 site or the CRE slightly reduced the promoter activity (60 or
90%, respectively). Moreover, cotransfection experiments showed that a trans-acting factor, CCAAT enhancer binding protein
(C/EBP
), which binds to both the NF-IL6 site and the CRE, increased the promoter activity of the PES-2 gene mainly through the CRE.
C/EBP
mRNA was rapidly induced by LPS. Collectively, these results suggest that transcription of the PES-2 gene in vascular endothelial
cells is regulated through combination of the NF-IL6 site and the CRE and that C/EBP
functions as one of the trans-acting factors.
Footnotes
-
↵* This work was supported in part by grants from the Ministry of Health and Welfare, by the Ministry of Education, Science, and Culture of Japan, and also by Special Coordination Funds for Promoting Science and Technology from the Science and Technology Agency of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) D64068[GenBank].
-
↵1 The abbreviations used are:
- PES
-
prostaglandin-endoperoxide synthase
- CRE
-
cAMP response element
- NF-IL6
-
nuclear factor for interleukin-6 expression
- NF-κB
-
nuclear factor κB
- bp
-
base pair(s)
- kb
-
kilobase pair(s)
- BAEC
-
bovine arterial endothelial cell(s)
- DMEM
-
Dulbecco's modified Eagle's medium
- LPS
-
lipopolysaccharide
- TPA
-
12-O-tetradecanoylphorbol-13-acetate
- DMEM-SF
-
serum-free and antibiotic-free DMEM
- C/EBP
-
CCAAT enhancer binding protein.
-
- Received May 25, 1995.
- Revision received August 8, 1995.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
