Rapid estrogen-induced phosphorylation of the SRC-3 coactivator occurs in an extranuclear complex containing estrogen receptor

Mol Cell Biol. 2005 Sep;25(18):8273-84. doi: 10.1128/MCB.25.18.8273-8284.2005.


SRC-3/AIB1/ACTR/pCIP/RAC3/TRAM1 is a primary transcriptional coregulator for estrogen receptor (ER). Six SRC-3 phosphorylation sites have been identified, and these can be induced by steroids, cytokines, and growth factors, involving multiple kinase signaling pathways. Using phosphospecific antibodies for six phosphorylation sites, we investigated the mechanisms involved in estradiol (E2)-induced SRC-3 phosphorylation and found that this occurs only when either activated estrogen receptor alpha (ERalpha) or activated ERbeta is present. Both the activation function 1 and the ligand binding domains of ERalpha are required for maximal induction. Mutations in the coactivator binding groove of the ERalpha ligand binding domain inhibit E2-stimulated SRC-3 phosphorylation, as do mutations in the nuclear receptor-interacting domain of SRC-3, suggesting that ERalpha must directly contact SRC-3 for this posttranslational modification to take place. A transcriptionally inactive ERalpha mutant which localizes to the cytoplasm supports E2-induced SRC-3 phosphorylation. Mutations of the ERalpha DNA binding domain did not block this rapid E2-dependent SRC-3 phosphorylation. Together these data demonstrate that E2-induced SRC-3 phosphorylation is dependent on a direct interaction between SRC-3 and ERalpha and can occur outside of the nucleus. Our results provide evidence for an early nongenomic action of ER on SRC-3 that supports the well-established downstream genomic roles of estrogen and coactivators.

Publication types

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

MeSH terms

  • Acetyltransferases / immunology
  • Acetyltransferases / metabolism*
  • Amino Acid Motifs
  • Antibodies, Phospho-Specific / immunology
  • Cell Nucleus / metabolism
  • Cytoplasm / chemistry
  • Cytoplasm / metabolism*
  • DNA / metabolism
  • Estradiol / pharmacology*
  • Estrogen Receptor alpha / analysis
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism*
  • Estrogen Receptor beta / analysis
  • Estrogen Receptor beta / genetics
  • Estrogen Receptor beta / metabolism*
  • Histone Acetyltransferases
  • Humans
  • Mutation
  • Nuclear Proteins / metabolism
  • Nuclear Receptor Coactivator 3
  • Oncogene Proteins / immunology
  • Oncogene Proteins / metabolism*
  • Phosphorylation / drug effects
  • Protein Structure, Tertiary
  • Receptors, Interferon / metabolism
  • Selective Estrogen Receptor Modulators / pharmacology
  • Trans-Activators / immunology
  • Trans-Activators / metabolism*


  • Antibodies, Phospho-Specific
  • Estrogen Receptor alpha
  • Estrogen Receptor beta
  • IFNGR2 protein, human
  • Nuclear Proteins
  • Oncogene Proteins
  • Receptors, Interferon
  • Selective Estrogen Receptor Modulators
  • Trans-Activators
  • Estradiol
  • DNA
  • Acetyltransferases
  • Histone Acetyltransferases
  • NCOA3 protein, human
  • Nuclear Receptor Coactivator 3