Reconstitution of mitotic chromatids with a minimum set of purified factors

Keishi Shintomi, Tatsuro Takahashi, Tatsuya Hirano

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

The assembly of mitotic chromosomes, each composed of a pair of rod-shaped chromatids, is an essential prerequisite for accurate transmission of the genome during cell division. It remains poorly understood, however, how this fundamental process might be achieved and regulated in the cell. Here we report an in vitro system in which mitotic chromatids can be reconstituted by mixing a simple substrate with only six purified factors: core histones, three histone chaperones (nucleoplasmin, Nap1 and FACT), topoisomerase II (topo II) and condensin I. We find that octameric nucleosomes containing the embryonic variant H2A.X-F are highly susceptible to FACT and function as the most productive substrate for subsequent actions of topo II and condensin I. Cdk1 phosphorylation of condensin I is the sole mitosis-specific modification required for chromatid reconstitution. This experimental system will enhance our understanding of the mechanisms of action of individual factors and their cooperation during this process.

Original languageEnglish
Pages (from-to)1014-1023
Number of pages10
JournalNature Cell Biology
Volume17
Issue number8
DOIs
Publication statusPublished - Aug 6 2015

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Chromatids
Type II DNA Topoisomerase
Type I DNA Topoisomerase
Nucleoplasmins
Histone Chaperones
Nucleosomes
Mitosis
Cell Division
Histones
Chromosomes
Phosphorylation
Genome
condensin complexes

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Reconstitution of mitotic chromatids with a minimum set of purified factors. / Shintomi, Keishi; Takahashi, Tatsuro; Hirano, Tatsuya.

In: Nature Cell Biology, Vol. 17, No. 8, 06.08.2015, p. 1014-1023.

Research output: Contribution to journalArticle

Shintomi, Keishi ; Takahashi, Tatsuro ; Hirano, Tatsuya. / Reconstitution of mitotic chromatids with a minimum set of purified factors. In: Nature Cell Biology. 2015 ; Vol. 17, No. 8. pp. 1014-1023.
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