Loss of full-length DNA replication regulator Rif1 in two-cell embryos is associated with zygotic transcriptional activation
| Autor: | Tomio Ono, Satoshi Yamazaki, Kaoru Mita-Yoshida, Yasumasa Nishito, Hisao Masai, Naoko Yoshizawa-Sugata |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
DNA Replication
Transcriptional Activation Zygote Telomere-Binding Proteins 4-OHT 4-hydroxytamoxifen IDR intrinsic disordered region Biochemistry 2C two-cell (embryo) Hpf hours post fertilization Histones Mice Transcription (biology) Animals Dox doxycycline Enhancer Molecular Biology Gene KD knockdown Transcriptional bursting Mice Inbred ICR KO knockout ERV endogenous retrovirus Chemistry ES embryonic stem DNA replication Acetylation Cell Biology DNA Methylation Chromatin Up-Regulation Cell biology DNA-Binding Proteins Mice Inbred C57BL IVF in vitro fertilization DNA methylation RT room temperature Ectopic expression Research Article Transcription Factors |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/j.jbc.2021.101367 |
| Popis: | Rif1 regulates DNA replication timing and double-strand break repair, and its depletion induces transcriptional bursting of 2-cell (2C) zygote-specific genes in mouse ES cells. However, how Rif1 regulates zygotic transcription is unclear. We show here that Rif1 depletion promotes the formation of a unique Zscan4 enhancer structure harboring both histone H3 lysine 27 acetylation (H3K27ac) and moderate levels of silencing chromatin mark H3K9me3. Curiously, another enhancer mark H3K4me1 is missing whereas DNA methylation is still maintained in the structure, which spreads across gene bodies and neighboring regions within the Zscan4 gene cluster. We also found by function analyses of Rif1 domains in ES cells that ectopic expression of Rif1 lacking N-terminal domain results in upregulation of 2C transcripts. This appears to be caused by dominant negative inhibition of endogenous Rif1 protein localization at the nuclear periphery through formation of hetero-oligomers between the N-terminally truncated and endogenous forms. Strikingly, in murine 2-cell embryos, most of Rif1-derived polypeptides are expressed as truncated forms in soluble nuclear or cytosolic fraction, and are likely non-functional. Toward the morula stage, the full-length form of Rif1 gradually increased. Our results suggest that the absence of the functional full-length Rif1 due to its instability or alternative splicing and potential inactivation of Rif1 through dominant inhibition by N-terminally truncated Rif1 polypeptides, may be involved in 2C-specific transcription program. |
| Databáze: | OpenAIRE |
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