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  A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis

Schellhaus, A., Magalska, A., Schooley, A., & Antonin, W. (2015). A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis. Journal of Visualized Experiments, 2015(106): e53407. doi:10.3791/53407.

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 Creators:
Schellhaus, AK1, Author                 
Magalska, A, Author                 
Schooley, A1, Author                 
Antonin, W1, Author                 
Affiliations:
1Antonin Group, Friedrich Miescher Laboratory, Max Planck Society, ou_3375712              

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 Abstract: During the vertebrate cell cycle chromatin undergoes extensive structural and functional changes. Upon mitotic entry, it massively condenses into rod shaped chromosomes which are moved individually by the mitotic spindle apparatus. Mitotic chromatin condensation yields chromosomes compacted fifty-fold denser as in interphase. During exit from mitosis, chromosomes have to re-establish their functional interphase state, which is enclosed by a nuclear envelope and is competent for replication and transcription. The decondensation process is morphologically well described, but in molecular terms poorly understood: We lack knowledge about the underlying molecular events and to a large extent the factors involved as well as their regulation. We describe here a cell-free system that faithfully recapitulates chromatin decondensation in vitro, based on mitotic chromatin clusters purified from synchronized HeLa cells and X. laevis egg extract. Our cell-free system provides an important tool for further molecular characterization of chromatin decondensation and its co-ordination with processes simultaneously occurring during mitotic exit such as nuclear envelope and pore complex re-assembly.

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 Dates: 2015-12
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.3791/53407
PMID: 26710245
 Degree: -

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Title: Journal of Visualized Experiments
  Other : Journal of Visualized Experiments: JoVE
  Abbreviation : J. Vis. Exp.
Source Genre: Journal
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Publ. Info: Rockville Pike, Bethesda MD : JoVE
Pages: 9 Volume / Issue: 2015 (106) Sequence Number: e53407 Start / End Page: - Identifier: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X