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In vivo protein‐DNA interactions at the kinin B 1 receptor gene promoter: No modification on interleukin‐1 beta or lipopolysaccharide induction
Author(s) -
Angers Martin,
Drouin Régen,
Bachvarova Magdalena,
Paradis Isabelle,
Marceau François,
Bachvarov Dimcho R.
Publication year - 2000
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(20000801)78:2<278::aid-jcb10>3.0.co;2-f
Subject(s) - microbiology and biotechnology , biology , reporter gene , promoter , gene , gene expression , transcriptional regulation , tata box , transcription (linguistics) , transcription factor , regulation of gene expression , genetics , linguistics , philosophy
The kinin B 1 receptor (B 1 R) gene is strongly upregulated following tissue injury and inflammation. In an attempt to define the regulatory elements that account for the control of B 1 R gene expression, we have conducted in vivo footprinting analysis of the B 1 R gene promoter region in three human cell types: embryonic lung fibroblast cells (IMR‐90), embryonic kidney cells (HEK‐293), and primary cultures of vascular umbilical smooth muscle cells. Initial in vitro delineation of the B 1 R gene promoter by transient transfection experiments with a reporter gene indicated that a 1.4‐kb region, located just upstream of the transcription initiation site, bears all the characteristics of a core promoter with a functional TATA box and additional positive and negative control elements, as some of them could be tissue‐specific. In vivo ultraviolet and dimethylsulfate footprinting analyses of the 1.4‐kb region revealed no difference between the footprint patterns in the three cell types studied. We found that even in the noninduced state, the B 1 R gene promoter is possibly bound by several sequence‐specific DNA binding proteins (GATA‐1, PEA3, AP‐1, CAAT, Sp1, Pit‐1a, Oct‐1, CREB). Some other footprints were detected on sequences that do not correspond to any known transcription factor binding site. No additional changes in protein‐DNA complexes were observed upon treatment with interleukin‐1 beta (IL‐1β) or bacterial lipopolysaccharide, shown previously to induce B 1 R gene expression. These results indicate that complex protein‐DNA interactions exist at the B 1 R gene promoter prior to induction by external stimuli even in cells (HEK‐293) that do not express a functional B 1 R. J. Cell. Biochem. 78:278–296, 2000. © 2000 Wiley‐Liss, Inc.