Methods for chromatin assembly and remodeling.
Packaging of the DNA in nucleosomes restricts its accessibility to regulatory factors and enzymatic complexes, making a local remodeling of the nucleosome structure a prerequisite to the establishment of protein-DNA interactions. The use of an experimental system in which one nucleosome is reconstituted on a short linear DNA fragment allows gel fractionation of nucleosomes according to their translational positions, whose locations are dependent on the underlying DNA sequence. Nucleosome mobilization by chromatin remodeling factors is easily detected by observing band disappearance in gel, which in turn provides evidence for histone octamer displacement. Here, we provide methods for chromatin assembly that we have been using in our analysis for nucleosome mobilization by chromatin remodeling factors. These methods are straightforward and easy to follow. Thus, they may provide a good starting assay system for analysis of nucleosome movements by other chromatin remodeling machines.
Duband-Goulet
Isabelle
I
Institut Andre Lwoff, CNRS UPR 9079, Equipe : fonction et dynamique de la chromatine, Bat B, 2ème Etage, 07 rue Guy Moquet, 94800 Villejuif, France.
Ouararhni
Khalid
K
Hamiche
Ali
A
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D004330:Drosophila; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008958:Models, Molecular; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism; D013395:Sucrose; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
C120419:Acf protein, Drosophila; D002843:Chromatin; D029721:Drosophila Proteins; D009707:Nucleosomes; D014157:Transcription Factors; D013395:Sucrose; D000255:Adenosine Triphosphate; D004247:DNA