Mapping the dimerization and DNA binding of nuclear receptors in live cells by a novel microscopic method

2020, October 13 - 10:45
Mapping the dimerization and DNA binding of nuclear receptors in live cells by a novel microscopic method

Bálint Rehó, Gábor Mocsár, Lina Fadel, Péter Brázda, László Nagy, György Vámosi and their colleagues from Germany, Lukas Lau, Gabriele Müller and Katalin Tóth developed together a microscopic method, and examined the dimerization and DNA-binding of nuclear receptors in live cells. The method (SPIM-FRET-FCCS) can be used to simultaneously map the proximity (using Förster resonance energy transfer, FRET), the joint motion and the mobility (using fluorescence cross-correlation spectroscopy, FCCS) of fluorescently tagged molecules in a selected plane of the cell, thus it is suitable for studying protein-protein and protein-DNA interactions. Retinoic acid receptor (RAR) binds to DNA and regulates the transcription of its target genes in a ligand-dependent manner, and serves as a therapeutic target in several diseases. It was demonstrated by the new method that agonist ligand enhances RAR dimerization with the retinoid X receptor and also increases the dimer’s DNA binding. The paper was published in the journal Analytical Chemistry doi: 10.1074/jbc.RA119.011614 (IF: 6.78).

 

 

Picture: Demonstrating the distinction of coexpressed protein pairs based on their proximity and co-mobility. Left: 2D maps of proximity (E: FRET efficiency) and co-mobility (rCCF: apparent co-diffusing fraction) measured from a GFP-mCherry fusion protein (GC+) connected by a short linker. Right: Cellular median values distinguishing high FRET, high comobility (GC+), low FRET high comobility (GC-: dyes connected by a long linker) and low FRET, low co-mobility (G/C: separately expressed dyes) populations.


Dear User!

 

The University of Debrecen considers the protection of personal data and the information provided to be of paramount importance. We hereby inform you that the University of Debrecen has reviewed its processes and incorporated the requirements of GDPR into his own data management and data protection activities, which entered mandatory on may 25,2018. The personal data of the users has been carefully handled by the University of Debrecen, it complies with the applicable data management regulations.Following the requirements of GDPR, we updated our Privacy Statement, which you can access by clicking on the link below: Privacy Policy.

I have read, understand and agree to terms.