Synergistic Regulation of Coregulator/Nuclear Receptor Interaction by Ligand and DNA

Ian Mitchelle S. de Vera; Jie Zheng; Scott Novick; Jinsai Shang; Travis S. Hughes; Richard Brust; Paola Munoz-Tello; William J. Gardner; David P. Marciano; Xiangming Kong; Patrick R. Griffin; Douglas J. Kojetin

doi:10.1016/j.str.2017.07.019

Synergistic Regulation of Coregulator/Nuclear Receptor Interaction by Ligand and DNA

Ian Mitchelle S. de Vera
, Jie Zheng
, Scott Novick
, Jinsai Shang
, Travis S. Hughes
, Richard Brust
, Paola Munoz-Tello
, William J. Gardner
, David P. Marciano
, Xiangming Kong
, Patrick R. Griffin
, Douglas J. Kojetin

Research output: Contribution to journal › Article › peer-review

46 Scopus citations

Abstract

Nuclear receptor (NR) transcription factors bind various coreceptors, small-molecule ligands, DNA response element sequences, and transcriptional coregulator proteins to affect gene transcription. Small-molecule ligands and DNA are known to influence receptor structure, coregulator protein interaction, and function; however, little is known on the mechanism of synergy between ligand and DNA. Using quantitative biochemical, biophysical, and solution structural methods, including ¹³C-detected nuclear magnetic resonance and hydrogen/deuterium exchange (HDX) mass spectrometry, we show that ligand and DNA cooperatively recruit the intrinsically disordered steroid receptor coactivator-2 (SRC-2/TIF2/GRIP1/NCoA-2) receptor interaction domain to peroxisome proliferator-activated receptor gamma-retinoid X receptor alpha (PPARγ-RXRα) heterodimer and reveal the binding determinants of the complex. Our data reveal a thermodynamic mechanism by which DNA binding propagates a conformational change in PPARγ-RXRα, stabilizes the receptor ligand binding domain dimer interface, and impacts ligand potency and cooperativity in NR coactivator recruitment. de Vera et al. combine biophysical and atomic-resolution solution structural methods to show that ligand and DNA binding to PPARγ-RXRα heterodimer cooperatively enhances recruitment of the SRC-2 coactivator, thereby revealing a complex allosteric communication pathway integrating signals from ligand, DNA, and coactivator.

Original language	English
Pages (from-to)	1506-1518.e4
Journal	Structure
Volume	25
Issue number	10
DOIs	https://doi.org/10.1016/j.str.2017.07.019
State	Published - Oct 3 2017

Funding

We thank Wolfgang Bermel (Bruker Biospin) for providing 13 C-detected NMR pulse sequences and advice, Mark Rance (University of Cincinnati) for a critical reading of the manuscript, and Natacha Rochel (IGBMC) for providing the elongated PPARγ-RXRα structural model. This work was supported in part by the James and Esther King Biomedical Research Program , Florida Department of Health ( 1KN-09 ); the William R. Kenan, Jr. Charitable Trust (TSRI High School Student Summer Internship Program); NIH grants R01DK101871 (D.J.K.), R01DK105825 (P.R.G.), F32DK097890 (T.H.), K99DK103116 (T.H.), and F32DK108442 (R.B.); and American Heart Association (AHA) fellowship awards 12POST12050025 (T.H.) and 16POST27780018 (R.B.).

Funders	Funder number
American Heart Association	12POST12050025, 16POST27780018
R01DK101871, F32DK108442, R01DK105825, F32DK097890
R00DK103116
Florida Department of Health	1KN-09

Keywords

NMR spectroscopy
allostery
cooperativity
hydrogen/deuterium exchange (HDX) mass spectrometry
ligand binding
nuclear receptor
peroxisome proliferator-activated receptor
retinoid X receptor
stabilization
transcription

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10.1016/j.str.2017.07.019

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@article{d89051d9a27240ec9545839cc2e1e259,

title = "Synergistic Regulation of Coregulator/Nuclear Receptor Interaction by Ligand and DNA",

abstract = "Nuclear receptor (NR) transcription factors bind various coreceptors, small-molecule ligands, DNA response element sequences, and transcriptional coregulator proteins to affect gene transcription. Small-molecule ligands and DNA are known to influence receptor structure, coregulator protein interaction, and function; however, little is known on the mechanism of synergy between ligand and DNA. Using quantitative biochemical, biophysical, and solution structural methods, including 13C-detected nuclear magnetic resonance and hydrogen/deuterium exchange (HDX) mass spectrometry, we show that ligand and DNA cooperatively recruit the intrinsically disordered steroid receptor coactivator-2 (SRC-2/TIF2/GRIP1/NCoA-2) receptor interaction domain to peroxisome proliferator-activated receptor gamma-retinoid X receptor alpha (PPARγ-RXRα) heterodimer and reveal the binding determinants of the complex. Our data reveal a thermodynamic mechanism by which DNA binding propagates a conformational change in PPARγ-RXRα, stabilizes the receptor ligand binding domain dimer interface, and impacts ligand potency and cooperativity in NR coactivator recruitment. de Vera et al. combine biophysical and atomic-resolution solution structural methods to show that ligand and DNA binding to PPARγ-RXRα heterodimer cooperatively enhances recruitment of the SRC-2 coactivator, thereby revealing a complex allosteric communication pathway integrating signals from ligand, DNA, and coactivator.",

keywords = "NMR spectroscopy, allostery, cooperativity, hydrogen/deuterium exchange (HDX) mass spectrometry, ligand binding, nuclear receptor, peroxisome proliferator-activated receptor, retinoid X receptor, stabilization, transcription",

author = "\{de Vera\}, \{Ian Mitchelle S.\} and Jie Zheng and Scott Novick and Jinsai Shang and Hughes, \{Travis S.\} and Richard Brust and Paola Munoz-Tello and Gardner, \{William J.\} and Marciano, \{David P.\} and Xiangming Kong and Griffin, \{Patrick R.\} and Kojetin, \{Douglas J.\}",

note = "Publisher Copyright: {\textcopyright} 2017 Elsevier Ltd",

year = "2017",

month = oct,

day = "3",

doi = "10.1016/j.str.2017.07.019",

language = "English",

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T1 - Synergistic Regulation of Coregulator/Nuclear Receptor Interaction by Ligand and DNA

AU - de Vera, Ian Mitchelle S.

AU - Zheng, Jie

AU - Novick, Scott

AU - Shang, Jinsai

AU - Hughes, Travis S.

AU - Brust, Richard

AU - Munoz-Tello, Paola

AU - Gardner, William J.

AU - Marciano, David P.

AU - Kong, Xiangming

AU - Griffin, Patrick R.

AU - Kojetin, Douglas J.

PY - 2017/10/3

Y1 - 2017/10/3

N2 - Nuclear receptor (NR) transcription factors bind various coreceptors, small-molecule ligands, DNA response element sequences, and transcriptional coregulator proteins to affect gene transcription. Small-molecule ligands and DNA are known to influence receptor structure, coregulator protein interaction, and function; however, little is known on the mechanism of synergy between ligand and DNA. Using quantitative biochemical, biophysical, and solution structural methods, including 13C-detected nuclear magnetic resonance and hydrogen/deuterium exchange (HDX) mass spectrometry, we show that ligand and DNA cooperatively recruit the intrinsically disordered steroid receptor coactivator-2 (SRC-2/TIF2/GRIP1/NCoA-2) receptor interaction domain to peroxisome proliferator-activated receptor gamma-retinoid X receptor alpha (PPARγ-RXRα) heterodimer and reveal the binding determinants of the complex. Our data reveal a thermodynamic mechanism by which DNA binding propagates a conformational change in PPARγ-RXRα, stabilizes the receptor ligand binding domain dimer interface, and impacts ligand potency and cooperativity in NR coactivator recruitment. de Vera et al. combine biophysical and atomic-resolution solution structural methods to show that ligand and DNA binding to PPARγ-RXRα heterodimer cooperatively enhances recruitment of the SRC-2 coactivator, thereby revealing a complex allosteric communication pathway integrating signals from ligand, DNA, and coactivator.

AB - Nuclear receptor (NR) transcription factors bind various coreceptors, small-molecule ligands, DNA response element sequences, and transcriptional coregulator proteins to affect gene transcription. Small-molecule ligands and DNA are known to influence receptor structure, coregulator protein interaction, and function; however, little is known on the mechanism of synergy between ligand and DNA. Using quantitative biochemical, biophysical, and solution structural methods, including 13C-detected nuclear magnetic resonance and hydrogen/deuterium exchange (HDX) mass spectrometry, we show that ligand and DNA cooperatively recruit the intrinsically disordered steroid receptor coactivator-2 (SRC-2/TIF2/GRIP1/NCoA-2) receptor interaction domain to peroxisome proliferator-activated receptor gamma-retinoid X receptor alpha (PPARγ-RXRα) heterodimer and reveal the binding determinants of the complex. Our data reveal a thermodynamic mechanism by which DNA binding propagates a conformational change in PPARγ-RXRα, stabilizes the receptor ligand binding domain dimer interface, and impacts ligand potency and cooperativity in NR coactivator recruitment. de Vera et al. combine biophysical and atomic-resolution solution structural methods to show that ligand and DNA binding to PPARγ-RXRα heterodimer cooperatively enhances recruitment of the SRC-2 coactivator, thereby revealing a complex allosteric communication pathway integrating signals from ligand, DNA, and coactivator.

KW - NMR spectroscopy

KW - allostery

KW - cooperativity

KW - hydrogen/deuterium exchange (HDX) mass spectrometry

KW - ligand binding

KW - nuclear receptor

KW - peroxisome proliferator-activated receptor

KW - retinoid X receptor

KW - stabilization

KW - transcription

UR - https://www.scopus.com/pages/publications/85030458846

U2 - 10.1016/j.str.2017.07.019

DO - 10.1016/j.str.2017.07.019

M3 - Article

C2 - 28890360

AN - SCOPUS:85030458846

SN - 0969-2126

VL - 25

SP - 1506-1518.e4

JO - Structure

JF - Structure

IS - 10

ER -

Synergistic Regulation of Coregulator/Nuclear Receptor Interaction by Ligand and DNA

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Keywords

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