Endothelial expression of endothelin-converting enzyme-1 beta mRNA is regulated by the transcription factor Ets-1.
Cleavage of big endothelins (ETs) by endothelin-converting enzymes (ECEs) represents the final step in the biosynthesis of ETs. ECE-1 is expressed predominantly in endothelial cells and exists in two isoforms, termed alpha and beta, differing in their 5' termini. We have recently shown that isoform-specific mRNA expression is directed by alternative promoters. To investigate possible mechanisms of transcriptional regulation of ECE-1, we stimulated E.A. hy 926 cells with phorbol ester and found a greater than threefold increase in ECE-1 beta mRNA at 12-24 h of stimulation. Because the beta-specific promoter is characterized by multiple consensus sequences for transcription factors of the ETS family, Ets-1 and PEA3, we also analyzed Ets-1 mRNA expression and found at least a fivefold increase in Ets-1 mRNA at 3 h of phorbol ester stimulation. Gel shift analysis revealed a specific interaction of nuclear proteins isolated from E.A. hy 926 cells with an oligonucleotide harboring the Ets-1 consensus sequence. Using a specific anti-Ets-1 antibody, we detected a supershifted band indicating the expression of Ets-1 protein in E.A. hy 926 cells. We conclude that Ets-1 is involved in transcriptional upregulation of ECE-1 beta mRNA in E.A. hy 926 cells induced by phorbol ester.
Orzechowski
H D
HD
Institute of Clinical Pharmacology and Toxicology, Benjamin Franklin Hospital, Free University of Berlin, Germany.
Günther
A
A
Menzel
S
S
Funke-Kaiser
H
H
Richter
M
M
Bohnemeier
H
H
Paul
M
M
D016282:Aspartic Acid Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D015152:Blotting, Northern; D004591:Electrophoresis, Polyacrylamide Gel; D000072396:Endothelin-Converting Enzymes; D004730:Endothelium, Vascular / Q000201:enzymology* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006801:Humans; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics; D010703:Phorbol Esters / Q000494:pharmacology; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology*; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D015854:Up-Regulation
C494041:ETS1 protein, human; D010703:Phorbol Esters; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D014157:Transcription Factors; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; D000072396:Endothelin-Converting Enzymes