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Sci Rep. 2017 Jun 16;7(1):3678. doi: 10.1038/s41598-017-03718-5.

Structural Characterization of the SMRT Corepressor Interacting with Histone Deacetylase 7.

Author information

1
European Molecular Biology Laboratory, Grenoble Outstation, 71 Avenue des Martyrs, CS90181, 38042, Grenoble Cedex 9, France.
2
Unit of Virus Host-Cell Interactions (UVHCI), University Grenoble Alpes, CNRS, EMBL, 38042, Grenoble, France.
3
Dunn School of Pathology, University of Oxford, South Parks Road, OX13 RE, Oxford, UK.
4
Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, 38044, Grenoble, France. robert.schneider@univ-lille1.fr.
5
University Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France. robert.schneider@univ-lille1.fr.
6
Institut de Biologie Structurale (IBS), CEA, CNRS, University Grenoble Alpes, 38044, Grenoble, France.
7
European Molecular Biology Laboratory, Grenoble Outstation, 71 Avenue des Martyrs, CS90181, 38042, Grenoble Cedex 9, France. darren.hart@ibs.fr.
8
Unit of Virus Host-Cell Interactions (UVHCI), University Grenoble Alpes, CNRS, EMBL, 38042, Grenoble, France. darren.hart@ibs.fr.

Abstract

The 2525 amino acid SMRT corepressor is an intrinsically disordered hub protein responsible for binding and coordinating the activities of multiple transcription factors and chromatin modifying enzymes. Here we have studied its interaction with HDAC7, a class IIa deacetylase that interacts with the corepressor complex together with the highly active class I deacetylase HDAC3. The binding site of class IIa deacetylases was previously mapped to an approximate 500 amino acid region of SMRT, with recent implication of short glycine-serine-isoleucine (GSI) containing motifs. In order to characterize the interaction in detail, we applied a random library screening approach within this region and obtained a range of stable, soluble SMRT fragments. In agreement with an absence of predicted structural domains, these were characterized as intrinsically disordered by NMR spectroscopy. We identified one of them, comprising residues 1255-1452, as interacting with HDAC7 with micromolar affinity. The binding site was mapped in detail by NMR and confirmed by truncation and alanine mutagenesis. Complementing this with mutational analysis of HDAC7, we show that HDAC7, via its surface zinc ion binding site, binds to a 28 residue stretch in SMRT comprising a GSI motif followed by an alpha helix.

PMID:
28623264
PMCID:
PMC5473869
DOI:
10.1038/s41598-017-03718-5
[Indexed for MEDLINE]
Free PMC Article

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