Reconstitution of mitotic chromatids with a minimum set of purified factors

Keishi Shintomi, Tatsuro S. Takahashi, Tatsuya Hirano

Research output: Contribution to journalArticlepeer-review

112 Citations (Scopus)


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
Issue number8
Publication statusPublished - Aug 6 2015
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Cell Biology


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