PUB-NChIP--"in vivo biotinylation" approach to study chromatin in proximity to a protein of interest

Muhammad Shoaib (1) , Arman Kulyyassov , Chloé Robin (1) , Kinga Winczura , Pavel Tarlykov , Emmanuelle Despas , Patricia L Kannouche (2) , Erlan Ramanculov , Marc Lipinski (1) , Vasily Ogryzko (1)

1 UMR8126 - Signalisation, noyaux et innovations en cancérologie
2 UMR 8200 - Stabilité Génétique et Oncogenèse

We have developed an approach termed PUB-NChIP (proximity utilizing biotinylation with native ChIP) to purify and study the protein composition of chromatin in proximity to a nuclear protein of interest. It is based on coexpression of (1) a protein of interest, fused with the bacterial biotin ligase BirA, together with (2) a histone fused to a biotin acceptor peptide (BAP), which is specifically biotinylated by BirA-fusion in the proximity of the protein of interest. Using the RAD18 protein as a model, we demonstrate that the RAD18-proximal chromatin is enriched in some H4 acetylated species. Moreover, the RAD18-proximal chromatin containing a replacement histone H2AZ has a different pattern of H4 acetylation. Finally, biotin pulse-chase experiments show that the H4 acetylation pattern starts to resemble the acetylation pattern of total H4 after the proximity of chromatin to RAD18 has been lost.

Domaines: Sciences du Vivant [q-bio]

Muhammad Shoaib, Arman Kulyyassov, Chloé Robin, Kinga Winczura, Pavel Tarlykov, et al.. PUB-NChIP--"in vivo biotinylation" approach to study chromatin in proximity to a protein of interest. Genome Research, 2013, 23 (2), pp.331-340. ⟨10.1101/gr.134874.111⟩. ⟨hal-02159970⟩