title: A defined in vitro system to study ATP-dependent remodeling of short chromatin fibers.
abstract: ATP-dependent remodeling factors regulate chromatin structure by catalyzing processes such as nucleosome repositioning or conformational changes of nucleosomes. Predominantly, their enzymatic properties have been investigated using mononucleosomal substrates. However, short nucleosomal arrays represent a much better mimic of the physiological chromatin context. Here, we provide a protocol for the enzyme-free reconstitution of regularly spaced nucleosomal arrays. We then explain how these arrays can serve as substrates to monitor ATP-dependent nucleosome movements and changes in the accessibility of nucleosomal DNA.
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, USA.
authors.forename: Verena K
authors.initials: VK
authors.lastname: Maier
authors.forename: Peter B
authors.initials: PB
authors.lastname: Becker
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting; D017931:DNA Primers / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D006657:Histones / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D009707:Nucleosomes / Q000378:metabolism; D011786:Quality Control; D011851:Radioactivity
chemical_list: D002843:Chromatin; D017931:DNA Primers; D006657:Histones; D009707:Nucleosomes; D000255:Adenosine Triphosphate; D004247:DNA; D008836:Micrococcal Nuclease