Human endothelin-converting enzyme-1β mRNA expression is regulated by an alternative promoter

被引:10
作者
Funke-Kaiser, H [1 ]
Orzechowski, HD [1 ]
Richter, A [1 ]
Paul, M [1 ]
机构
[1] Free Univ Berlin, Univ Hosp Benjamin Franklin, Inst Clin Pharmacol & Toxicol, D-12200 Berlin, Germany
关键词
big endothelin; endothelin-converting enzymes; ECE-1; isoforms; luciferase reporter vector;
D O I
10.1097/00005344-199800001-00004
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The central step in endothelin biosynthesis is site-specific cleavage of big endothelins by endothelin-converting enzymes (ECEs). ECE-1 is a membrane-bound metalloprotease, predominantly but not exclusively expressed in endothelial cells. ECE-1 is expressed in two mRNA isoforms, termed a. and beta, which differ only in the 5'-terminal regions but are functionally very similar when expressed in vitro. The structure of the human ECE-1 gene suggests either alternative splicing or alternative promoters as underlying mechanisms of mRNA isoform expression. We have previously shown that the alpha-upstream region exerts promoter activity in endothelial cells. To clarify whether the 5'-untranslated region upstream of exon 3, which contains the beta-specific sequence, acts as an alternative transcriptional promoter, we sequenced and cloned 1,206 bp upstream of the beta-specific translation initiation codon in a luciferase reporter vector. After transfection, we detected strong promoter activity in primary cultured endothelial cells (HU-VECs, BAECs) but only marginal activity in the endothelial cell line ECV304 and in CHO cells. Maximal promoter activity was observed with the full-length construct, 1206 (136% of the SV40 promoter activity in BAECs). Transfection of serial deletion mutants indicated at least three major regulatory regions within the promoter. Our results are consistent with cell type-restricted action of the beta-promoter and, in conjunction with the previously reported transcriptional start sites, clearly prove the existence of an alternative beta-specific promoter located in intron 2 of the human ECE-1 gene.
引用
收藏
页码:S7 / S9
页数:3
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