title: Sodium butyrate-mediated Sp3 acetylation represses human insulin-like growth factor binding protein-3 expression in intestinal epithelial cells.
abstract: OBJECTIVE
  Butyrate concentrations in the gastrointestinal tract vary greatly with age. In intestinal epithelial cells, butyrate enhances gene transcription by increasing histone acetylation, rendering the nucleosome open to transcription factors. However, it inhibits human insulin-like growth factor binding protein (hIGFBP)-3 expression. We therefore hypothesized that butyrate also acts by regulating transcription factor acetylation.
  
  
  METHODS
  Gene regulation was examined in Caco-2 cells. RNA stability was measured after interruption of transcription. The activity of deletion mutations of the hIGFBP-3 promoter was examined in reporter assays. Transcription factor binding to promoter DNA was analyzed.
  
  
  RESULTS
  Butyrate did not increase the transcription of a repressor because it inhibited hIGFBP-3 mRNA in the absence of protein synthesis. Nor did butyrate decrease the stability of hIGFBP-3 mRNA. Analysis of the hIGFBP-3 promoter demonstrated a butyrate-response element that included the binding sites for p300 and Sp1/Sp3. Transfection of Caco-2 cells with E1A, an inhibitor of p300 acetyltransferase activity, reversed the butyrate-induced repression of hIGFBP-3. Because Sp3 represses the initiation of transcription, we studied whether butyrate induced Sp3 acetylation. Electrophoretic mobility shift assays of nuclei extracted from Caco-2 cells treated with 5 mmol/L butyrate demonstrated an extra, heavier band in addition to the Sp3-DNA binding in untreated cells. This corresponded to a protein, detected only in butyrate treated cells, that was identified both by an anti-Sp3 antibody and by an anti-acetyl lysine antibody.
  
  
  CONCLUSIONS
  This study demonstrates that butyrate increases the acetylation of a nonhistone protein, Sp3, catalyzed by p300 acetyltransferase activity.
authors.affiliation: Centre for Adult and Paediatric Gastroenterology, Institute of Cell and Molecular Science, Barts and The London, Queen Mary School of Medicine and Dentistry, University of London, London E1 2AD, United Kingdom.
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: White
authors.forename: Peter
authors.initials: P
authors.lastname: Mulligan
authors.forename: Peter J
authors.initials: PJ
authors.lastname: King
authors.forename: Ian R
authors.initials: IR
authors.lastname: Sanderson
mesh_terms: D000107:Acetylation / Q000187:drug effects*; D002087:Butyrates / Q000494:pharmacology*; D018938:Caco-2 Cells; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D004847:Epithelial Cells / Q000378:metabolism; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051705:Sp3 Transcription Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor
chemical_list: D002087:Butyrates; D018797:Cell Cycle Proteins; D051548:Histone Acetyltransferases; D018972:Insulin-Like Growth Factor Binding Protein 3; D012313:RNA; D051705:Sp3 Transcription Factor; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C497121:SP3 protein, human