A multiprotein binding interface in an intrinsically disordered region of the tumor suppressor protein interferon regulatory factor-1.

The interferon-regulated transcription factor and tumor suppressor protein IRF-1 is predicted to be largely disordered outside of the DNA-binding domain. One of the advantages of intrinsically disordered protein domains is thought to be their ability to take part in multiple, specific but low affinity protein interactions; however, relatively few IRF-1-interacting ...
proteins have been described. The recent identification of a functional binding interface for the E3-ubiquitin ligase CHIP within the major disordered domain of IRF-1 led us to ask whether this region might be employed more widely by regulators of IRF-1 function. Here we describe the use of peptide aptamer-based affinity chromatography coupled with mass spectrometry to define a multiprotein binding interface on IRF-1 (Mf2 domain; amino acids 106-140) and to identify Mf2-binding proteins from A375 cells. Based on their function as known transcriptional regulators, a selection of the Mf2 domain-binding proteins (NPM1, TRIM28, and YB-1) have been validated using in vitro and cell-based assays. Interestingly, although NPM1, TRIM28, and YB-1 all bind to the Mf2 domain, they have differing amino acid specificities, demonstrating the degree of combinatorial diversity and specificity available through linear interaction motifs.
Mesh Terms:
Amino Acid Motifs, Amino Acid Sequence, Cell Line, Tumor, Chromatography, Affinity, DNA-Binding Proteins, Gene Expression Regulation, Humans, Interferon Regulatory Factor-1, Molecular Sequence Data, Nuclear Proteins, Peptides, Protein Binding, Protein Interaction Mapping, Repressor Proteins, Sequence Homology, Amino Acid, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Ubiquitin-Protein Ligases, Y-Box-Binding Protein 1
J. Biol. Chem.
Date: Apr. 22, 2011
Download Curated Data For This Publication
139916
Switch View:
  • Interactions 6