Tracking H3K27me3 and H4K20me1 during XCI reveals similarities in enrichment dynamics
Tjalsma, S. J. D.; Hori, M.; Sato, Y.; Bousard, A.; Ohi, A.; Raposo, A. C.; Roensch, J.; Le Saux, A.; Nogami, J.; Maehara, K.; Kujirai, T.; Handa, T.; Bages-Arnal, S.; Ohkawa, Y.; Kurumizaka, H.; da Rocha, S. T.; Zylicz, J. J.; Kimura, H.; Heard, E.
Show abstract
During X chromosome inactivation (XCI), in female placental mammals, gene silencing is initiated by the Xist long-noncoding 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 molecular mechanism allowing for H4K20me1 enrichment. To follow XCI dynamics in living cells, we developed a genetically-encoded, H3K27me3-specific intracellular antibody, or H3K27me3-mintbody. By combining it with live-imaging of H4K20me1, the X chromosome and Xist RNA we uncover similarities in the initial accumulation dynamics of H3K27me3 and H4K20me1. Further ChIP-seq analysis confirmed concurrent accumulation of both marks during XCI albeit with distinct genomic distributions. Using a Xist B and C repeat mutant, which can silence the X but does not allow for H3K27me3 deposition, we also found a lack of H4K20me1 enrichment. Thus, these two marks accumulate at the X thanks to the same region of Xist and H4K20me1 in particular may have a role in the chromatin compaction that characterises facultative heterochromatin.
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