Remodeling and Repositioning of Nucleosomes in Nucleosomal Arrays.
| Autor: | Ludwigsen J; Molecular Biology, Biomedical Center, Faculty of Medicine, LMU Munich, Martinsried, 82152, Germany.; Proteros Biostructures GmbH, Bunsenstraße 7a, Martinsried, 82152, Germany., Hepp N; Molecular Biology, Biomedical Center, Faculty of Medicine, LMU Munich, Martinsried, 82152, Germany.; Department of Endocrinology, Copenhagen University Hospital Hvidovre, Kettegaards Allé 30, Hvidovre, 2650, Denmark., Klinker H; Molecular Biology, Biomedical Center, Faculty of Medicine, LMU Munich, Martinsried, 82152, Germany.; Biomedical Center, LMU Munich, Martinsried, 82152, Germany., Pfennig S; Molecular Biology, Biomedical Center, Faculty of Medicine, LMU Munich, Martinsried, 82152, Germany.; Clariant Produkte (Deutschland) GmbH, Semmelweisstraße 1, Planegg, 82152, Germany., Mueller-Planitz F; Molecular Biology, Biomedical Center, Faculty of Medicine, LMU Munich, Martinsried, 82152, Germany. fmueller@bmc.med.lmu.de. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1805, pp. 349-370. |
| DOI: | 10.1007/978-1-4939-8556-2_18 |
| Abstrakt: | ATP-dependent nucleosome remodeling factors sculpt the nucleosomal landscape of eukaryotic chromatin. They deposit or evict nucleosomes or reposition them along DNA in a process termed nucleosome sliding. Remodeling has traditionally been analyzed using mononucleosomes as a model substrate. In vivo, however, nucleosomes form extended arrays with regular spacing. Here, we describe how regularly spaced nucleosome arrays can be reconstituted in vitro and how these arrays can be used to dissect remodeling in the test tube. We outline two assays. The first assay senses various structural changes to a specific nucleosome within the nucleosomal array whereas the second assay is specific toward detecting repositioning of nucleosomes within the array. Both assays exploit changes to the accessibility of DNA to restriction enzymes during the remodeling reaction. |
| Databáze: | MEDLINE |
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