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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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26710245 ???displayArticle.pmcLink???PMC4694036 ???displayArticle.link???J Vis Exp
Belmont,
Mitotic chromosome structure and condensation.
2006, Pubmed
Belmont,
Mitotic chromosome structure and condensation.
2006,
Pubmed Bürglin,
Cloning of nucleoplasmin from Xenopus laevis oocytes and analysis of its developmental expression.
1987,
Pubmed
,
Xenbase Chan,
In vitro study of nuclear assembly and nuclear import using Xenopus egg extracts.
2006,
Pubmed
,
Xenbase de la Barre,
Assembly of mitotic chromosomes in Xenopus egg extract.
1999,
Pubmed
,
Xenbase Eisenhardt,
Xenopus in vitro assays to analyze the function of transmembrane nucleoporins and targeting of inner nuclear membrane proteins.
2014,
Pubmed
,
Xenbase Forbes,
Spontaneous formation of nucleus-like structures around bacteriophage DNA microinjected into Xenopus eggs.
1983,
Pubmed
,
Xenbase Galy,
A role for gp210 in mitotic nuclear-envelope breakdown.
2008,
Pubmed Gant,
Nuclear assembly.
1997,
Pubmed Gasser,
Improved methods for the isolation of individual and clustered mitotic chromosomes.
1987,
Pubmed Gillespie,
Preparation and use of Xenopus egg extracts to study DNA replication and chromatin associated proteins.
2012,
Pubmed
,
Xenbase Hartl,
Nuclear assembly with lambda DNA in fractionated Xenopus egg extracts: an unexpected role for glycogen in formation of a higher order chromatin intermediate.
1994,
Pubmed
,
Xenbase Lavoie,
Mitotic chromosome condensation requires Brn1p, the yeast homologue of Barren.
2000,
Pubmed
,
Xenbase Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase Magalska,
RuvB-like ATPases function in chromatin decondensation at the end of mitosis.
2014,
Pubmed
,
Xenbase Maresca,
Methods for studying spindle assembly and chromosome condensation in Xenopus egg extracts.
2006,
Pubmed
,
Xenbase Paulson,
Isolation of chromosome clusters from metaphase-arrested HeLa cells.
1982,
Pubmed Philpott,
Sperm decondensation in Xenopus egg cytoplasm is mediated by nucleoplasmin.
1991,
Pubmed
,
Xenbase Philpott,
Nucleoplasmin remodels sperm chromatin in Xenopus egg extracts.
1992,
Pubmed
,
Xenbase Ramadan,
Cdc48/p97 promotes reformation of the nucleus by extracting the kinase Aurora B from chromatin.
2007,
Pubmed
,
Xenbase Sandaltzopoulos,
Transcriptional repression by nucleosomes but not H1 in reconstituted preblastoderm Drosophila chromatin.
1994,
Pubmed Ulbert,
Direct membrane protein-DNA interactions required early in nuclear envelope assembly.
2006,
Pubmed
,
Xenbase Whitaker,
Calcium at fertilization and in early development.
2006,
Pubmed
,
Xenbase Wignall,
The condensin complex is required for proper spindle assembly and chromosome segregation in Xenopus egg extracts.
2003,
Pubmed
,
Xenbase Zhang,
Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
