In vivo DNase I-mediated footprinting analysis along the human bradykinin B1 receptor (BDKRB1) gene promoter: evidence for cell-specific regulation

Biochem J. 2005 Jul 1;389(Pt 1):37-46. doi: 10.1042/BJ20042104.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Cells, Cultured
  • DNA Footprinting*
  • Deoxyribonuclease I / metabolism*
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / genetics*
  • Humans
  • Interleukin-1 / pharmacology
  • Lymphocytes / metabolism
  • Molecular Sequence Data
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism
  • Myocytes, Smooth Muscle / metabolism
  • Nucleosomes / drug effects
  • Nucleosomes / metabolism
  • Organ Specificity
  • Promoter Regions, Genetic / genetics*
  • Receptor, Bradykinin B1 / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Serum
  • Transcription Factor TFIIB / metabolism
  • YY1 Transcription Factor / metabolism

Substances

  • Interleukin-1
  • Nucleosomes
  • Receptor, Bradykinin B1
  • Transcription Factor TFIIB
  • YY1 Transcription Factor
  • YY1 protein, human
  • Deoxyribonuclease I