Interference mapping of nuclear protein binding to the acute phase response element of the mouse α1 acid glycoprotein gene

Jeffrey P. Rabek, John Papaconstantinou

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Exchange in binding of transcription factors C/EBPα and C/EBPβ at a regulatory site in the α1-acid glycoprotein (AGP) promoter, termed the acute phase response element (APRE), has been correlated with bacterial lipopolysaccharide (LPS) mediated induction. The APRE contains overlapping recognition sequences for C/EBP's and glucocorticoid receptor (GR). Electrophortetic mobility shift assays show that this site can bind both GR and C/EBP. However, using liver nuclear extract, which contains GR binding activity, only C/EBP binds to the APRE. Binding interference methods, using dimethyl sulfate and potassium permanganate modification of specific bases, detected interference only with modification of bases that are in the region of the C/EBP binding site that do not overlap with the GRE sequence. There are no significant differences between the interference patterns of control and LPS treated liver nuclear extracts, suggesting that the region of close contact between protein and DNA is similar for C/EBPα (untreated) and C/EBPβ (treated).

Original languageEnglish (US)
Pages (from-to)608-613
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume255
Issue number3
DOIs
StatePublished - Feb 24 1999

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Acute-Phase Reaction
Glucocorticoid Receptors
Response Elements
Nuclear Proteins
Protein Binding
Liver Extracts
Glycoproteins
Genes
Liver
Acids
Lipopolysaccharides
Potassium Permanganate
Electrophoretic Mobility Shift Assay
Assays
Transcription Factors
Binding Sites
DNA
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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abstract = "Exchange in binding of transcription factors C/EBPα and C/EBPβ at a regulatory site in the α1-acid glycoprotein (AGP) promoter, termed the acute phase response element (APRE), has been correlated with bacterial lipopolysaccharide (LPS) mediated induction. The APRE contains overlapping recognition sequences for C/EBP's and glucocorticoid receptor (GR). Electrophortetic mobility shift assays show that this site can bind both GR and C/EBP. However, using liver nuclear extract, which contains GR binding activity, only C/EBP binds to the APRE. Binding interference methods, using dimethyl sulfate and potassium permanganate modification of specific bases, detected interference only with modification of bases that are in the region of the C/EBP binding site that do not overlap with the GRE sequence. There are no significant differences between the interference patterns of control and LPS treated liver nuclear extracts, suggesting that the region of close contact between protein and DNA is similar for C/EBPα (untreated) and C/EBPβ (treated).",
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N2 - Exchange in binding of transcription factors C/EBPα and C/EBPβ at a regulatory site in the α1-acid glycoprotein (AGP) promoter, termed the acute phase response element (APRE), has been correlated with bacterial lipopolysaccharide (LPS) mediated induction. The APRE contains overlapping recognition sequences for C/EBP's and glucocorticoid receptor (GR). Electrophortetic mobility shift assays show that this site can bind both GR and C/EBP. However, using liver nuclear extract, which contains GR binding activity, only C/EBP binds to the APRE. Binding interference methods, using dimethyl sulfate and potassium permanganate modification of specific bases, detected interference only with modification of bases that are in the region of the C/EBP binding site that do not overlap with the GRE sequence. There are no significant differences between the interference patterns of control and LPS treated liver nuclear extracts, suggesting that the region of close contact between protein and DNA is similar for C/EBPα (untreated) and C/EBPβ (treated).

AB - Exchange in binding of transcription factors C/EBPα and C/EBPβ at a regulatory site in the α1-acid glycoprotein (AGP) promoter, termed the acute phase response element (APRE), has been correlated with bacterial lipopolysaccharide (LPS) mediated induction. The APRE contains overlapping recognition sequences for C/EBP's and glucocorticoid receptor (GR). Electrophortetic mobility shift assays show that this site can bind both GR and C/EBP. However, using liver nuclear extract, which contains GR binding activity, only C/EBP binds to the APRE. Binding interference methods, using dimethyl sulfate and potassium permanganate modification of specific bases, detected interference only with modification of bases that are in the region of the C/EBP binding site that do not overlap with the GRE sequence. There are no significant differences between the interference patterns of control and LPS treated liver nuclear extracts, suggesting that the region of close contact between protein and DNA is similar for C/EBPα (untreated) and C/EBPβ (treated).

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