The Biophysical Basis for Phosphorylation-Enhanced DNA-Binding Autoinhibition of the ETS1 Transcription Factor

JOURNAL OF MOLECULAR BIOLOGY

卷:431

Article

Perez-Borrajero, Cecilia^1,2  Lin, Chang Sheng-Huei³  Okon, Mark^2,4  Scheu, Karlton²  Graves, Barbara J.^5,6  Murphy, Michael E. P.³  McIntosh, Lawrence P.^1,2,4,7 

[1] Univ British Columbia, Genome Sci & Technol Program, Vancouver, BC V6T 1Z3, Canada

[2] Univ British Columbia, Dept Biochem & Mol Biol, Vancouver, BC V6T 1Z3, Canada

[3] Univ British Columbia, Dept Microbiol & Immunol, Vancouver, BC V6T 1Z3, Canada

[4] Univ British Columbia, Dept Chem, Vancouver, BC V6T 1Z1, Canada

[5] Univ Utah, Huntsman Canc Inst, Sch Med, Dept Oncol Sci, Salt Lake City, UT 84112 USA

[6] Howard Hughes Med Inst, Chevy Chase, MD 20815 USA

[7] Univ British Columbia, Michael Smith Labs, Vancouver, BC V6T 1Z4, Canada

关键词: intrinsically disordered regions;    fuzzy complexes;    protein structure and dynamics;    NMR spectroscopy;    X-ray crystallography;   

DOI  :  10.1016/j.jmb.2018.12.011

来源: Elsevier

【 摘 要 】

The eukaryotic transcription factor ETS1 is regulated by an intrinsically disordered serine-rich region (SRR) that transiently associates with the adjacent ETS domain to inhibit DNA binding. In this study, we further elucidated the physicochemical basis for ETS1 autoinhibition by characterizing the interaction of its ETS domain with a series of synthetic peptides corresponding to the SRR. Binding is driven by the hydrophobic effect and enhanced electrostatically by phosphorylation of serines adjacent to aromatic residues in the amphipathic SRR. Structural characterization of the dynamic peptide/protein complex by NMR spectroscopy and X-ray crystallography revealed multiple modes of binding that lead to autoinhibition by synergistically blocking the DNA-binding interface of the ETS domain and stabilizing an appended helical inhibitory module against allosterically induced unfolding. Consistent with these conclusions, the SRR peptide does not interact with DNA-bound ETS1. In addition, we found that the ETS1 SRR phosphopeptide binds to distantly related PU.1 in vitro, indicating that autoinhibition exploits features of the ETS domain that are conserved across this family of transcription factors. (C) 2018 Elsevier Ltd. All rights reserved.

【 授权许可】

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