Ras/mitogen-activated protein kinase signaling activates Ets-1 and Ets-2 by CBP/p300 recruitment.

Cell signaling affects gene expression by regulating the activity of transcription factors. Here, we report that mitogen-activated protein kinase (MAPK) phosphorylation of Ets-1 and Ets-2, at a conserved site N terminal to their Pointed (PNT) domains, resulted in enhanced transactivation by preferential recruitment of the coactivators CREB binding protein (CBP) ...
and p300. We discovered this phosphorylation-augmented interaction in an unbiased affinity chromatography screen of HeLa nuclear extracts by using either mock-treated or ERK2-phosphorylated ETS proteins as ligands. Binding between purified proteins demonstrated a direct interaction. Both the phosphoacceptor site, which lies in an unstructured region, and the PNT domain were required for the interaction. Minimal regions that were competent for induced CBP/p300 binding in vitro also supported MAPK-enhanced transcription in vivo. CBP coexpression potentiated MEK1-stimulated Ets-2 transactivation of promoters with Ras-responsive elements. Furthermore, CBP and Ets-2 interacted in a phosphorylation-enhanced manner in vivo. This study describes a distinctive interface for a transcription factor-coactivator complex and demonstrates a functional role for inducible CBP/p300 binding. In addition, our findings decipher the mechanistic link between Ras/MAPK signaling and two specific transcription factors that are relevant to both normal development and tumorigenesis.
Mesh Terms:
Acetyltransferases, Amino Acid Sequence, Baculoviridae, Blotting, Western, CREB-Binding Protein, Cell Cycle Proteins, Chromatography, Affinity, Genes, Reporter, Genes, ras, Hela Cells, Histone Acetyltransferases, Humans, Luciferases, Mitogen-Activated Protein Kinases, Models, Biological, Models, Molecular, Nuclear Magnetic Resonance, Biomolecular, Nuclear Proteins, Phosphorylation, Protein Structure, Tertiary, Proto-Oncogene Protein c-ets-1, Proto-Oncogene Protein c-ets-2, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-ets, Recombinant Proteins, Signal Transduction, Silver Staining, Trans-Activators, Transcription Factors, Transcriptional Activation, p300-CBP Transcription Factors
Mol. Cell. Biol.
Date: Dec. 01, 2004
Download Curated Data For This Publication
Switch View:
  • Interactions 9