The prereplication complex recruits XEco2 to chromatin to promote cohesin acetylation in Xenopus egg extracts

Torahiko L. Higashi, Megumi Ikeda, Hiroshi Tanaka, Takuro Nakagawa, Masashige Bando, Katsuhiko Shirahige, Yumiko Kubota, Haruhiko Takisawa, Hisao Masukata, Tatsuro Takahashi

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Background: Sister chromatids are held together by the ring-shaped cohesin complex, which is loaded onto chromosomes before DNA replication. Cohesion between sister chromosomes is established during DNA replication, and it requires acetylation of the Smc3 subunit of cohesin by evolutionally conserved cohesin acetyltransferases (CoATs). However, how CoATs are recruited to chromatin and how cohesin acetylation is regulated remain unclear. Results: We found that cohesin acetylation requires pre-RC-dependent chromatin loading of cohesin, but surprisingly, it is independent of DNA synthesis in Xenopus egg extracts. Immunodepletion experiments revealed that XEco2 is the CoAT responsible for Smc3 acetylation and sister chromatid cohesion. Recruitment of XEco2 onto chromatin was dependent on pre-RC assembly but was independent of cohesin loading and DNA synthesis. Two short N-terminal motifs, PBM-A and PBM-B, which are conserved among vertebrate Esco2/XEco2 homologs, were collectively essential for pre-RC-dependent chromatin association of XEco2, cohesin acetylation, and subsequent sister chromatid cohesion. The conserved PCNA-interacting protein box in XEco2 was largely dispensable for Smc3 acetylation but was partially required for cohesion. Interaction of acetylated cohesin with DNA was stabilized against salt-wash treatments after DNA replication. Conclusions: Our results demonstrate that pre-RC formation regulates chromatin association of XEco2 in Xenopus egg extracts. We propose that this reaction is critical to acetylate cohesin, whose DNA binding is subsequently stabilized by DNA replication.

Original languageEnglish
Pages (from-to)977-988
Number of pages12
JournalCurrent Biology
Volume22
Issue number11
DOIs
Publication statusPublished - Jun 5 2012
Externally publishedYes

Fingerprint

Acetylation
Xenopus
acetylation
Chromatin
Ovum
chromatin
DNA replication
cohesion
acetyltransferases
chromatids
extracts
DNA
DNA Replication
Acetyltransferases
Chromatids
chromosomes
synthesis
Chromosomes
cohesins
vertebrates

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

The prereplication complex recruits XEco2 to chromatin to promote cohesin acetylation in Xenopus egg extracts. / Higashi, Torahiko L.; Ikeda, Megumi; Tanaka, Hiroshi; Nakagawa, Takuro; Bando, Masashige; Shirahige, Katsuhiko; Kubota, Yumiko; Takisawa, Haruhiko; Masukata, Hisao; Takahashi, Tatsuro.

In: Current Biology, Vol. 22, No. 11, 05.06.2012, p. 977-988.

Research output: Contribution to journalArticle

Higashi, TL, Ikeda, M, Tanaka, H, Nakagawa, T, Bando, M, Shirahige, K, Kubota, Y, Takisawa, H, Masukata, H & Takahashi, T 2012, 'The prereplication complex recruits XEco2 to chromatin to promote cohesin acetylation in Xenopus egg extracts', Current Biology, vol. 22, no. 11, pp. 977-988. https://doi.org/10.1016/j.cub.2012.04.013
Higashi, Torahiko L. ; Ikeda, Megumi ; Tanaka, Hiroshi ; Nakagawa, Takuro ; Bando, Masashige ; Shirahige, Katsuhiko ; Kubota, Yumiko ; Takisawa, Haruhiko ; Masukata, Hisao ; Takahashi, Tatsuro. / The prereplication complex recruits XEco2 to chromatin to promote cohesin acetylation in Xenopus egg extracts. In: Current Biology. 2012 ; Vol. 22, No. 11. pp. 977-988.
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AU - Bando, Masashige

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