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Upregulation of human heme oxygenase gene expression by Ets‐family proteins
Author(s) -
Deramaudt Bertrand M.J.M.,
Remy P.,
Abraham N.G.
Publication year - 1999
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19990301)72:3<311::aid-jcb1>3.0.co;2-g
Subject(s) - heme oxygenase , chloramphenicol acetyltransferase , electrophoretic mobility shift assay , angiogenesis , transcription factor , microbiology and biotechnology , heme , biology , gene expression , promoter , downregulation and upregulation , signal transduction , gene , chemistry , biochemistry , genetics , enzyme
Overexpression of human heme oxygenase‐1 has been shown to have the potential to promote EC proliferation and angiogenesis. Since Ets‐family proteins have been shown to play an important role in angiogenesis, we investigated the presence of ETS binding sites (EBS), GGAA/T, and ETS protein contributing to human HO‐1 gene expression. Several chloramphenicol acetyltransferase constructs were examined in order to analyze the effect of ETS family proteins on the transduction of HO‐1 in Xenopus oocytes and in microvessel endothelial cells. Heme oxygenase promoter activity was up‐regulated by FLI‐1ERGETS‐1 protein(s). Chloramphenicol acetyltransferase (CAT) assays demonstrated that the promoter region (−1500 to +19) contains positive and negative control elements and that all three members of the ETS protein family were responsible for the up‐regulation of HHO‐1. Electrophoretic mobility shift assays (EMSA), performed with nuclear extracts from endothelial cells overexpressing HHO‐1 gene, and specific HHO‐1 oligonucleotides probes containing putative EBS resulted in a specific and marked bandshift. Synergistic binding was observed in EMSA between AP‐1 on the one hand, FLI‐1, ERG, and ETS‐1 protein on the other. Moreover, 5′‐deletion analysis demonstrated the existence of a negative control element of HHO‐1 expression located between positions −1500 and −120 on the HHO‐1 promoter. The presence of regulatory sequences for transcription factors such as ETS‐1, FLI‐1, or ERG, whose activity is associated with cell proliferation, endothelial cell differentiation, and matrix metalloproteinase transduction, may be an indication of the important role that HO‐1 may play in coronary collateral circulation, tumor growth, angiogenesis, and hemoglobin‐induced endothelial cell injuries. J. Cell. Biochem. 72:311–321, 1999. © 1999 Wiley‐Liss, Inc.