title: Purification of a specific native genomic locus for proteomic analysis.
abstract: Here, we describe an approach to isolate native chromatin sections without genomic engineering for label-free proteomic identification of associated proteins and histone post-translational modifications. A transcription activator-like (TAL) protein A fusion protein was designed to recognize a unique site in the yeast GAL1 promoter. The TAL-PrA fusion enabled chromatin affinity purification (ChAP) of a small section of native chromatin upstream from the GAL1 locus, permitting mass spectrometric (MS) identification of proteins and histone post-translational modifications regulating galactose-induced transcription. This TAL-ChAP-MS approach allows the biochemical isolation of a specific native genomic locus for proteomic studies and will provide for unprecedented objective insight into protein and epigenetic mechanisms regulating site-specific chromosome metabolism.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA and Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Stephanie D
authors.initials: SD
authors.lastname: Byrum
authors.forename: Sean D
authors.initials: SD
authors.lastname: Taverna
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Tackett
mesh_terms: D002843:Chromatin / Q000302:isolation & purification*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005686:Galactokinase / Q000235:genetics; D056426:Genetic Loci; D023281:Genomics; D006657:Histones / Q000378:metabolism*; D013058:Mass Spectrometry; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; D029701:Saccharomyces cerevisiae Proteins; C524219:GAL1 protein, S cerevisiae; D005686:Galactokinase
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