The structural and dynamic basis of Ets-1 DNA binding autoinhibition

Gregory M Lee; Logan W Donaldson; Miles A Pufall; Hyun-Seo Kang; Isabelle Pot; Barbara J Graves; Lawrence P McIntosh

doi:10.1074/jbc.M410722200

Back

The structural and dynamic basis of Ets-1 DNA binding autoinhibition

Journal article

Open access

Peer reviewed

The structural and dynamic basis of Ets-1 DNA binding autoinhibition

Gregory M Lee, Logan W Donaldson, Miles A Pufall, Hyun-Seo Kang, Isabelle Pot, Barbara J Graves and Lawrence P McIntosh

The Journal of biological chemistry, Vol.280(8), pp.7088-7099

02/25/2005

DOI: 10.1074/jbc.M410722200

PMID: 15591056

Files and links (1)

url

https://doi.org/10.1074/jbc.M410722200View

Published (Version of record) Open Access

Abstract

The transcription factor Ets-1 is regulated by the allosteric coupling of DNA binding with the unfolding of an alpha-helix (HI-1) within an autoinhibitory module. To understand the structural and dynamic basis for this autoinhibition, we have used NMR spectroscopy to characterize Ets-1DeltaN301, a partially inhibited fragment of Ets-1. The NMR-derived Ets-1DeltaN301 structure reveals that the autoinhibitory module is formed predominantly by the hydrophobic packing of helices from the N-terminal (HI-1, HI-2) and C-terminal (H4, H5) inhibitory sequences, along with H1 of the intervening DNA binding ETS domain. The intramolecular interactions made by HI-1 in Ets-1DeltaN301 are similar to the intermolecular contacts observed in the crystal structure of an Ets-1DeltaN300 dimer, confirming that the latter represents a domain-swapped species. (15)N relaxation studies demonstrate that the backbone of the N-terminal inhibitory sequence is mobile on the nanosecond-picosecond and millisecond-microsecond time scales. Furthermore, hydrogen exchange measurements reveal that amide protons in helices HI-1 and HI-2 exchange with water at rates only approximately 15- and approximately 75-fold slower, respectively, than predicted for an unfolded polypeptide. These findings indicate that inhibitory helices are only marginally stable even in the absence of DNA. The energetic coupling of DNA binding with the facile unfolding of the labile HI-1 provides a mechanism for modulating Ets-1 DNA binding activity via protein partnerships, post-translational modifications, or mutations. Ets-1 autoinhibition illustrates how conformational equilibria within structural domains can regulate macromolecular interactions.

Protein Structure, Tertiary

Proto-Oncogene Proteins - metabolism

Allosteric Regulation

Models, Molecular

DNA - metabolism

Transcription Factors - metabolism

Deuterium Exchange Measurement

Peptide Fragments - chemistry

Animals

Hydrophobic and Hydrophilic Interactions

Nuclear Magnetic Resonance, Biomolecular

Mice

Proto-Oncogene Protein c-ets-1

Proto-Oncogene Proteins c-ets

Details

Title: Subtitle: The structural and dynamic basis of Ets-1 DNA binding autoinhibition
Creators: Gregory M Lee - Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada
Logan W Donaldson
Miles A Pufall
Hyun-Seo Kang
Isabelle Pot
Barbara J Graves
Lawrence P McIntosh
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.280(8), pp.7088-7099
DOI: 10.1074/jbc.M410722200
PMID: 15591056
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM38663 / NIGMS NIH HHS R01 GM038663 / NIGMS NIH HHS CA93247 / NCI NIH HHS CA24014 / NCI NIH HHS
Language: English
Date published: 02/25/2005
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984024558202771

Metrics

9 Record Views

85 Times Cited - Web of Science