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J Biol Chem. 2019 Feb 8;294(6):1904-1914. doi: 10.1074/jbc.RA118.005550. Epub 2018 Dec 14.

A versatile mouse model of epitope-tagged histone H3.3 to study epigenome dynamics.

Author information

1
From the Division of Developmental Biology and.
2
the Department of Immunology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
3
the Section on Molecular Endocrinology, NICHD, National Institutes of Health, Bethesda, Maryland 20892 and.
4
From the Division of Developmental Biology and ozatok@nih.gov.

Abstract

The variant histone H3.3 is incorporated into the genome in a transcription-dependent manner. This histone is thus thought to play a role in epigenetic regulation. However, our understanding of how H3.3 controls gene expression and epigenome landscape has remained incomplete. This is partly because precise localization of H3.3 in the genome has been difficult to decipher particularly for cells in vivo To circumvent this difficulty, we generated knockin mice, by homologous recombination, to replace both of the two H3.3 loci (H3f3a and H3f3b) with the hemagglutinin-tagged H3.3 cDNA cassette, which also contained a GFP gene. We show here that the hemagglutinin-tagged H3.3 and GFP are expressed in the majority of cells in all adult tissues tested. ChIP-seq data, combined with RNA-seq, revealed a striking correlation between the level of transcripts and that of H3.3 accumulation in expressed genes. Finally, we demonstrate that H3.3 deposition is markedly enhanced upon stimulation by interferon on interferon-stimulated genes, highlighting transcription-coupled H3.3 dynamics. Together, these H3.3 knockin mice serve as a useful experimental model to study epigenome regulation in development and in various adult cells in vivo.

KEYWORDS:

ChIP-sequencing (ChIP-seq); H3.3; Histone variant; chromatin; epigenetics; interferon; transcription

PMID:
30552116
PMCID:
PMC6369307
[Available on 2020-02-08]
DOI:
10.1074/jbc.RA118.005550
[Indexed for MEDLINE]

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