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Biophys Chem. 2016 Mar;210:2-8. doi: 10.1016/j.bpc.2015.10.006. Epub 2015 Oct 30.

Monitoring of the retinoic acid receptor-retinoid X receptor dimerization upon DNA binding by native mass spectrometry.

Author information

1
Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UMR 7140 CNRS/Université de Strasbourg - "Chimie de la Matière Complexe", 1 Rue Blaise Pascal, 67008 Strasbourg, France.
2
Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Institut National de Santé et de Recherche Médicale (INSERM) U964, Centre National de Recherche Scientifique (CNRS) UMR 7104, Université de Strasbourg, 1 rue Laurent Fries, 67404 Illkirch, France.
3
Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UMR 7140 CNRS/Université de Strasbourg - "Chimie de la Matière Complexe", 1 Rue Blaise Pascal, 67008 Strasbourg, France. Electronic address: npotier@unistra.fr.

Abstract

Identifying protein-DNA interactions is essential to understand the regulatory networks of cells and their influence on gene expression. In this study, we use native electrospray mass spectrometry (ESI-MS) to investigate how the heterodimerization of retinoic acid receptor-retinoid X receptor (RAR-RXR) is mediated by DNA sequence. In presence of various RAR response elements (RAREs), three oligomeric states of RAR-RXR DNA binding domains (DBDs) bound to RAREs (monomer, homo- or heterodimers) were detected and individually monitored to follow subunit assembly and disassembly upon RAREs' abundancy or sequence. In particular, a cooperative heterodimerization was shown with RARb2 DR5 (5 base pair spaced direct repeat) while a high heterogeneity reflecting random complex formation could be observed with the DR0 response elements, in agreement with native gel electrophoresis data or molecular modeling. Such MS information will help to identify the composition of species formed in solution and to define which DR sequence is specific for RAR-RXR heterodimerization.

KEYWORDS:

Dynamics study; Native mass spectrometry; Nuclear receptors; Protein–DNA interaction; RAR–RXR complexes; Retinoic acid receptor (RAR); Retinoid X receptor (RXR)

PMID:
26558701
DOI:
10.1016/j.bpc.2015.10.006
[Indexed for MEDLINE]

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