Document Detail

PUB-NChIP--"in vivo biotinylation" approach to study chromatin in proximity to a protein of interest.
MedLine Citation:
PMID:  23038767     Owner:  NLM     Status:  MEDLINE    
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.
Muhammad Shoaib; Arman Kulyyassov; Chloé Robin; Kinga Winczura; Pavel Tarlykov; Emmanuelle Despas; Patricia Kannouche; Erlan Ramanculov; Marc Lipinski; Vasily Ogryzko
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-10-04
Journal Detail:
Title:  Genome research     Volume:  23     ISSN:  1549-5469     ISO Abbreviation:  Genome Res.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-04     Completed Date:  2013-07-15     Revised Date:  2013-08-12    
Medline Journal Info:
Nlm Unique ID:  9518021     Medline TA:  Genome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  331-40     Citation Subset:  IM    
UMR8126, Université Paris-Sud 11, CNRS, Institut de Cancérologie Gustave Roussy, 94805 Villejuif, France.
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MeSH Terms
Biotinylation / methods
Cell Line
Chromatin / metabolism*
Chromatin Immunoprecipitation / methods*
Nuclear Proteins / metabolism*
Protein Processing, Post-Translational
Reg. No./Substance:
0/Chromatin; 0/Histones; 0/Nuclear Proteins

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