Remodeling of Individual Nucleosomes in Nucleosome Arrays

Publikation: Beitrag in Buch/Konferenzbericht/Sammelband/GutachtenBeitrag in Buch/Sammelband/GutachtenBeigetragenBegutachtung

Beitragende

  • Petra Vizjak - , Institut für Physiologische Chemie, Ludwig-Maximilians-Universität München (LMU), Boehringer Ingelheim GmbH (Autor:in)
  • Nicola Hepp - , Copenhagen University Hospitals, Medizinische Fakultät Carl Gustav Carus Dresden, Institut für Physiologische Chemie (Autor:in)
  • Felix Mueller-Planitz - , Institut für Physiologische Chemie (Autor:in)

Abstract

Adenosin triphosphate (ATP)-dependent nucleosome remodeling factors sculpt the nucleosomal landscape of eukaryotic chromatin. They deposit, evict, or reposition nucleosomes 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. Both assays exploit the changes in the accessibility of DNA to restriction enzymes during the remodeling reaction. The first assay uses the restriction enzyme to cleave the restriction site as soon as it becomes accessible during remodeling. As such, this assay mostly reports the kinetic parameter of the "forward" reaction of nucleosome remodeling. In contrast, the second assay measures how fast a particular nucleosome in the array reaches its steady-state position.

Details

OriginalspracheEnglisch
TitelMolecular Motors
Redakteure/-innenChristophe Lavelle, Antoine Le Gall
ErscheinungsortNew York
Herausgeber (Verlag)Humana New York
Seiten271-291
Seitenumfang21
ISBN (elektronisch)978-1-0716-4280-1
ISBN (Print)978-1-0716-4279-5
PublikationsstatusVeröffentlicht - 2025
Peer-Review-StatusJa

Publikationsreihe

ReiheMethods in Molecular Biology (MIMB)
Band2881
ISSN1064-3745

Externe IDs

Scopus 85213538055

Schlagworte

ASJC Scopus Sachgebiete

Schlagwörter

  • Adenosin triphosphate (ATP)-dependent nucleosome remodeling, Chromatin reconstitution, Chromatin remodeling enzyme, Histone; Chromatin accessibility, ISWI, Nucleosome arrays, Nucleosome sliding/repositioning, Snf2-family ATPase