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Technology Development for Studying the Dynamics of Protein‐protein Associations on Chromatin
Author(s) -
Tackett Alan
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb155
Subject(s) - chromatin , histone , chromatin remodeling , histone modifying enzymes , histone code , computational biology , microbiology and biotechnology , biology , nucleosome , genetics , dna
We are optimizing technologies for quantitative analysis of proteins and histone post‐translational modifications associated with distinct chromatin loci. Key to employing these approaches is the sufficient use of chemical cross‐linking to minimize protein exchange during chromatin isolation; conversely, over cross‐linking renders chromatin insoluble. We developed a quantitative method termed transient I‐DIRT, useful for determining optimal levels of chromatin cross‐linking. We have applied transient I‐DIRT to define the local interactome of a low‐abundance yeast histone acetyltransferase (NuA3), and have also applied this method to determine cross‐linking conditions for efficient chromatin purification with minimal histone exchange. Our results provide insight into the dynamics of protein‐protein association on chromatin and establish the parameters for purification of native chromatin sections.