H4K20me1 and H3K27me3 are concurrently loaded onto the inactive X chromosome but dispensable for inducing gene silencing

Sjoerd J.D. Tjalsma, Mayako Hori, Yuko Sato, Aurelie Bousard, Akito Ohi, Ana Cláudia Raposo, Julia Roensch, Agnes Le Saux, Jumpei Nogami, Kazumitsu Maehara, Tomoya Kujirai, Tetsuya Handa, Sandra Bagés-Arnal, Yasuyuki Ohkawa, Hitoshi Kurumizaka, Simão Teixeira da Rocha, Jan J. Żylicz, Hiroshi Kimura, Edith Heard

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)


During X chromosome inactivation (XCI), in female placental mammals, gene silencing is initiated by the Xist long non-coding RNA. Xist accumulation at the X leads to enrichment of specific chromatin marks, including PRC2-dependent H3K27me3 and SETD8-dependent H4K20me1. However, the dynamics of this process in relation to Xist RNA accumulation remains unknown as is the involvement of H4K20me1 in initiating gene silencing. To follow XCI dynamics in living cells, we developed a genetically encoded, H3K27me3-specific intracellular antibody or H3K27me3-mintbody. By combining live-cell imaging of H3K27me3, H4K20me1, the X chromosome and Xist RNA, with ChIP-seq analysis we uncover concurrent accumulation of both marks during XCI, albeit with distinct genomic distributions. Furthermore, using a Xist B and C repeat mutant, which still shows gene silencing on the X but not H3K27me3 deposition, we also find a complete lack of H4K20me1 enrichment. This demonstrates that H4K20me1 is dispensable for the initiation of gene silencing, although it may have a role in the chromatin compaction that characterises facultative heterochromatin.

Original languageEnglish
Article numbere51989
JournalEMBO Reports
Issue number3
Publication statusPublished - Mar 3 2021

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Genetics


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