In vivo DNase I-mediated footprinting analysis along the human bradykinin B1 receptor (BDKRB1) gene promoter: evidence for cell-specific regulation.
By applying in vivo dimethyl sulphate and UV light type C-footprinting analysis, we previously showed that specific DNA sequences in the -1349/+42 core promoter region of the inducible human BDKRB1 (bradykinin B1 receptor) gene correlated with its transcriptional activity. In the present study we used the highly sensitive DNase I in vivo footprinting approach to delineate more precisely the functional domains of the BDKRB1 gene promoter in human SMCs (smooth muscle cells). Human lymphocytes that do not express a functional BDKRB1 were also studied as a reference using dimethyl sulphate, UV light type C and DNase I treatments. An obvious difference was found in the DNase I-footprinting patterns between cellular systems that express a functional BDKRB1 (SMCs) in comparison with human lymphocytes, where randomly distributed nucleosome-like footprinting patterns were found in the bulk of the core promoter region studied. Gel-shift assays and expression studies pointed to the implication of the YY1 and a TBP/TFIIB (TATA-box-binding protein/transcription factor IIB) transcription factor in the regulation of BDKRB1 gene expression in SMCs and possible YY1 involvement in the mechanisms of nuclear factor kappaB-mediated regulation of the receptor expression. No significant changes in the promoter foot-printing pattern were found after treatment with interleukin-1beta or serum (known BDKRB1 gene inducers), indicating that definite regulatory motifs could exist outside the BDKRB1 gene core promoter region studied.
Angers
Martin
M
Unité de Recherche en Génétique Humaine et Moléculaire, Research Centre, Hôpital St-François d'Assise, Centre Hospitalier Universitaire de Québec, 10 de l'Espinay Street, QC, G1L 3L5, Canada.
Drouin
Régen
R
Bachvarova
Magdalena
M
Paradis
Isabelle
I
Bissell
Brad
B
Hiromura
Makoto
M
Usheva
Anny
A
Bachvarov
Dimcho
D
D001483:Base Sequence; D002478:Cells, Cultured; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D008214:Lymphocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics*; D043783:Receptor, Bradykinin B1 / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum; D035581:Transcription Factor TFIIB / Q000378:metabolism; D050986:YY1 Transcription Factor / Q000378:metabolism
D007375:Interleukin-1; D009707:Nucleosomes; D043783:Receptor, Bradykinin B1; D035581:Transcription Factor TFIIB; D050986:YY1 Transcription Factor; C494698:YY1 protein, human; D003850:Deoxyribonuclease I