title: A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis.
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.
authors.affiliation: Friedrich Miescher Laboratory, Max Planck Society.
authors.forename: Anna K
authors.initials: AK
authors.lastname: Schellhaus
authors.affiliation: Nencki Institute of Experimental Biology, Polish Academy of Sciences.
authors.forename: Adriana
authors.initials: A
authors.lastname: Magalska
authors.affiliation: Friedrich Miescher Laboratory, Max Planck Society.
authors.forename: Allana
authors.initials: A
authors.lastname: Schooley
authors.affiliation: Friedrich Miescher Laboratory, Max Planck Society; wolfram.antonin@tuebingen.mpg.de.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Antonin
mesh_terms: D000818:Animals; D002474:Cell-Free System; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007399:Interphase / Q000502:physiology; D008938:Mitosis / Q000502:physiology*; D009685:Nuclear Envelope / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D002843:Chromatin