The STAT3 NH2-terminal domain stabilizes enhanceosome assembly by interacting with the p300 bromodomain.
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 NH(2) terminus-deficient STAT3 mutant (STAT3-DeltaN) was unable to efficiently induce either STAT3-mediated reporter activity or endogenous mRNA expression. Chromatin immunoprecipitation assays were performed to determine whether the NH(2)-terminal domain regulates p300 recruitment or stabilizes enhanceosome assembly. Despite equivalent levels of STAT3 binding, cells expressing the STAT3-DeltaN 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 NH(2)-terminal domain regulates its interaction with p300. In addition, the STAT3 NH(2)-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-DeltaB) exhibited a weaker binding to STAT3, and the enhanceosome formation on the socs3 promoter was inhibited when p300-DeltaB was overexpressed. Taken together, our data suggest that the STAT3 NH(2)-terminal domain plays an important role in the interleukin-6 signaling pathway by interacting with the p300 bromodomain, thereby stabilizing enhanceosome assembly.
Mesh Terms:
Amino Acid Substitution, Animals, Cell Line, Tumor, Embryo, Mammalian, Fibroblasts, Histone Acetyltransferases, Humans, Interleukin-6, Mice, Mice, Knockout, Mutation, Missense, Protein Binding, Protein Structure, Tertiary, RNA Polymerase II, STAT3 Transcription Factor, Signal Transduction, Transcription, Genetic, p300-CBP Transcription Factors
Amino Acid Substitution, Animals, Cell Line, Tumor, Embryo, Mammalian, Fibroblasts, Histone Acetyltransferases, Humans, Interleukin-6, Mice, Mice, Knockout, Mutation, Missense, Protein Binding, Protein Structure, Tertiary, RNA Polymerase II, STAT3 Transcription Factor, Signal Transduction, Transcription, Genetic, p300-CBP Transcription Factors
J. Biol. Chem.
Date: Nov. 07, 2008
PubMed ID: 18782771
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