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,Jan J. Żylicz,Jan J. Żylicz,Hiroshi Kimura,Edith Heard,Edith Heard +21 more
TLDR
In this article, a genetically encoded, H3K27me3-specific intracellular antibody or H4K20me1-mintbody was developed to follow X chromosome inactivation (XCI) dynamics in living cells.Abstract:
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.read more
Citations
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Single-cell profiling of transcriptome and histone modifications with EpiDamID
TL;DR: In this paper , DamID is used to detect histone post-translational modifications and transcription at the single-cell resolution, which can be used to profile singlecell Polycomb occupancy in mouse embryoid bodies and provide evidence for hierarchical gene regulatory networks.
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The X chromosome in C. elegans sex determination and dosage compensation.
TL;DR: In this article , the interplay between chromatin modification and three-dimensional chromosome structure imposed by an X-specific condensin complex to regulate gene expression over vast chromosomal territories was revealed.
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Mechanisms of sex determination and X-chromosome dosage compensation
TL;DR: This review analyzes the chromosome counting mechanism that tallies X-chromosome number to determine sex in the nematode Caenorhabditis elegans and the associated dosage compensation mechanism that balances X- chromatin modification and chromosome structure in regulating gene expression over vast chromosomal territories.
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Live-cell imaging probes to track chromatin modification dynamics.
TL;DR: Live-cell chromatin modification imaging using probes to visualize chromatin and its modifications will address dynamic chromatin regulation and will be useful for assaying and screening effective epigenome drugs in cells and organisms.
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Live imaging of transcription sites using an elongating RNA polymerase II-specific probe.
Satoshi Uchino,Yuma Ito,Yuko Sato,Tetsuya Handa,Yasuyuki Ohkawa,Makio Tokunaga,Hiroshi Kimura +6 more
TL;DR: In this paper, a modified Intacellular Antibilinear Antibody (mintbody) was used to detect the sites of RNAP2 Ser2ph-mintbody foci.
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