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Region Hovedstaden - en del af Københavns Universitetshospital
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Remodeling and Repositioning of Nucleosomes in Nucleosomal Arrays

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  • Johanna Ludwigsen
  • Nicola Hepp
  • Henrike Klinker
  • Sabrina Pfennig
  • Felix Mueller-Planitz
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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.

OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind1805
Sider (fra-til)349-370
ISSN1064-3745
DOI
StatusUdgivet - 2018

ID: 54888663