Binding of NF-κB to Nucleosomes: Effect of Translational Positioning, Nucleosome Remodeling and Linker Histone H1

Imtiaz Nisar Lone 1 Manu Shubhdarshan Shukla 1, 2 John Lalith Charles Richard 3 Zahary Jordanov Peshev 1, 4 Stefan Dimitrov 5 Dimitar Angelov 1
1 LBMC - Laboratoire de Biologie Moléculaire de la Cellule
2 Institut Albert Bonniot
3 Institut Albert Bonniot de Grenoble
LBMC - Laboratoire de Biologie Moléculaire de la Cellule, Institut Albert Bonniot
4 Institute of Electronics [Sofia]
5 INSERM U823 - Institut d'oncologie/développement Albert Bonniot de Grenoble
Abstract : NF-κB is a key transcription factor regulating the expression of inflammatory responsive genes. How NF-κB binds to naked DNA templates is well documented, but how it interacts with chromatin is far from being clear. Here we used a combination of UV laser footprinting, hydroxyl footprinting and electrophoretic mobility shift assay to investigate the binding of NF-κB to nucleosomal templates. We show that NF-κB p50 homodimer is able to bind to its recognition sequence, when it is localized at the edge of the core particle, but not when the recognition sequence is at the interior of the nucleosome. Remodeling of the nucleosome by the chromatin remodeling machine RSC was not sufficient to allow binding of NF-κB to its recognition sequence located in the vicinity of the nucleosome dyad, but RSC-induced histone octamer sliding allowed clearly detectable binding of NF-κB with the slid particle. Importantly, nucleosome dilution-driven removal of H2A-H2B dimer led to complete accessibility of the site located close to the dyad to NF-κB. Finally, we found that NF-κB was able to displace histone H1 and prevent its binding to nucleosome. These data provide important insight on the role of chromatin structure in the regulation of transcription of NF-κB dependent genes.
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Journal articles
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https://hal-ens-lyon.archives-ouvertes.fr/ensl-00961484
Contributor : Agnès Ganivet <>
Submitted on : Thursday, March 20, 2014 - 10:33:57 AM
Last modification on : Friday, September 6, 2019 - 11:14:03 AM

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  • HAL Id : ensl-00961484, version 1
  • DOI : 10.1371

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ENS-LYON | UGA | U823 | UNIV-LYON1

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Imtiaz Nisar Lone, Manu Shubhdarshan Shukla, John Lalith Charles Richard, Zahary Jordanov Peshev, Stefan Dimitrov, et al.. Binding of NF-κB to Nucleosomes: Effect of Translational Positioning, Nucleosome Remodeling and Linker Histone H1. PLoS Genetics, Public Library of Science, 2013, 9 (9), pp.e1003830. ⟨10.1371⟩. ⟨ensl-00961484⟩

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