Acetylation of Estrogen Receptor Alpha by p300 at Lysines 266 and 268 Enhances the Deoxyribonucleic Acid Binding and Transactivation Activities of the Receptor

Mol Endocrinol. 2006 Jul;20(7):1479-93. doi: 10.1210/me.2005-0531. Epub 2006 Feb 23.

Abstract

Using a variety of biochemical and cell-based approaches, we show that estrogen receptor alpha (ERalpha) is acetylated by the p300 acetylase in a ligand- and steroid receptor coactivator-dependent manner. Using mutagenesis and mass spectrometry, we identified two conserved lysine residues in ERalpha (Lys266 and Lys268) that are the primary targets of p300-mediated acetylation. These residues are acetylated in cells, as determined by immunoprecipitation-Western blotting experiments using an antibody that specifically recognizes ERalpha acetylated at Lys266 and Lys268. The acetylation of ERalpha by p300 is reversed by native cellular deacetylases, including trichostatin A-sensitive enzymes (i.e. class I and II deacetylases) and nicotinamide adenine dinucleotide-dependent/nicotinamide-sensitive enzymes (i.e. class III deacetylases, such as sirtuin 1). Acetylation at Lys266 and Lys268, or substitution of the same residues with glutamine (i.e. K266/268Q), a residue that mimics acetylated lysine, enhances the DNA binding activity of ERalpha in EMSAs. Likewise, substitution of Lys266 and Lys268 with glutamine enhances the ligand-dependent activity of ERalpha in a cell-based reporter gene assay. Collectively, our results implicate acetylation as a modulator of the ligand-dependent gene regulatory activity of ERalpha. Such regulation is likely to play a role in estrogen-dependent signaling outcomes in a variety of estrogen target tissues in both normal and pathological states.

Publication types

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

MeSH terms

  • Acetylation
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cats
  • Cell Compartmentation
  • Cells, Cultured
  • Conserved Sequence
  • Cricetinae
  • DNA / metabolism*
  • Estrogen Receptor alpha / metabolism*
  • Estrogen Receptor beta / metabolism
  • Estrogens / metabolism
  • Humans
  • Hydroxamic Acids / pharmacology
  • Lysine / metabolism*
  • Mass Spectrometry
  • Mice
  • Molecular Sequence Data
  • Niacinamide / pharmacology
  • Nuclear Receptor Coactivator 2 / metabolism
  • Point Mutation
  • Protein Structure, Tertiary
  • Rats
  • Sequence Alignment
  • Sirtuins / metabolism
  • Transcriptional Activation
  • p300-CBP Transcription Factors / metabolism*

Substances

  • Estrogen Receptor alpha
  • Estrogen Receptor beta
  • Estrogens
  • Hydroxamic Acids
  • Nuclear Receptor Coactivator 2
  • Niacinamide
  • trichostatin A
  • DNA
  • p300-CBP Transcription Factors
  • Sirtuins
  • Lysine