The STAT3 NH2-terminal domain stabilizes enhanceosome assembly by interacting with the p300 bromodomain

Tieying Hou, Sutapa Ray, Chang Lee, Allan R. Brasier

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Abstract

Signal transducer and activator of transcription 3 (STAT3) is a latent transcription factor mainly activated by the interleukin-6 cytokine family. Previous studies have shown that activated STAT3 recruits p300, a coactivator whose intrinsic histone acetyltransferase activity is essential for transcription. Here we investigated the function of the STAT3 NH 2-terminal domain and how its interaction with p300 regulates STAT3 signal transduction. In STAT3-/- mouse embryonic fibroblasts, a stably expressed NH2 terminus-deficient STAT3 mutant (STAT3-ΔN) was unable to efficiently induce either STAT3-mediated reporter activity or endogenous mRNA expression. Chromatin immunoprecipitation assays were performed to determine whether the NH2-terminal domain regulates p300 recruitment or stabilizes enhanceosome assembly. Despite equivalent levels of STAT3 binding, cells expressing the STAT3-ΔN mutant were unable to recruit p300 and RNA polymerase II to the native socs3 promoter as efficiently as those expressing STAT3-full length. We previously reported that the STAT3 NH 2-terminal domain is acetylated by p300 at Lys-49 and Lys-87. By introducing K49R/K87R mutations, here we found that the acetylation status of the STAT3 NH2-terminal domain regulates its interaction with p300. In addition, the STAT3 NH2-terminal binding site maps to the p300 bromodomain, a region spanning from amino acids 995 to 1255. Finally a p300 mutant lacking the bromodomain (p300-ΔB) exhibited a weaker binding to STAT3, and the enhanceosome formation on the socs3 promoter was inhibited when p300-ΔB was overexpressed. Taken together, our data suggest that the STAT3 NH2-terminal domain plays an important role in the interleukin-6 signaling pathway by interacting with the p300 bromodomain, thereby stabilizing enhanceosome assembly.

Original languageEnglish (US)
Pages (from-to)30725-30734
Number of pages10
JournalJournal of Biological Chemistry
Volume283
Issue number45
DOIs
StatePublished - Nov 7 2008

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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