Characterization of the c-specific promoter of the gene encoding human endothelin-converting enzyme-1 (ECE-1)
作者： H. Funke-Kaiser , J. Bolbrinker , S. Theis , J. Lemmer , C.-M. Richter
DOI: 10.1016/S0014-5793(00)01086-3
关键词:
摘要: Abstract Human ECE-1 is expressed in four isoforms with different tissue distribution and its mRNA protein levels are altered under certain pathophysiological conditions. To investigate the transcriptional regulation of ECE-1, we studied regulatory region ECE-1c, major isoform. A genomic clone comprising complete human gene including putative ECE-1c-specific promoter was obtained. Up to 968 bp upstream c-specific translation initiation start codon several serial deletion mutants were subcloned into a reporter vector transfected endothelial (BAEC, EA.hy926, ECV304) epithelial (MDA MB435S, MCF7) cells, showing very strong activity comparison SV40 previously described ECE-1a 1b promoters. Transfection indicated two positive regions within (−142/−240 −240/490) likely involved binding GATA ETS transcription factors. RNase protection assay (RPA) 5′-RACE revealed multiple sites located at about −110, −140 −350 bp. Site-directed mutagenesis demonstrated crucial role for E2F cis-element basal ECE-1c activity. Additionally, found correlation between isoform-specific corresponding ECE-1a, 1b, 1c activities.
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