Abstract
The Cys2/His2-type zinc finger protein Zfp296 has been implicated in stem cell pluripotency and tumor pathogenesis. However, its mechanisms remain elusive. Here, we demonstrated that a Zfp296 deficiency in mice impairs germ-cell development and embryonic growth. Zfp296 was intracellularly localized to heterochromatin in embryos. A GST-Zfp296 pull-down experiment using ES cell nuclear extract followed by LC-MS/MS showed that Zfp296 interacts with component proteins of heterochromatin (such as HP1, Dnmt1, Dnmt3b, and ATRX) and the NuRD complex. We focused on H3K9 methylation as a hallmark of heterochromatin, and found that Zfp296 overexpression in cultured cells reduces the Suv39h1-mediated H3K9 methylation. Consistent with this finding, in Zfp296 -/- mouse embryos, we observed a global increase in H3K9 methylation in a developmental stage-dependent manner, and showed, by ChIP-qPCR, that the H3K9me3 levels at major satellite repeats were elevated in Zfp296 -/- embryos. Our results demonstrate that Zfp296 is a component of heterochromatin that affects embryonic development by negatively regulating H3K9 methylation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Differentiation
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Chromobox Protein Homolog 5
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Chromosomal Proteins, Non-Histone / genetics
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Chromosomal Proteins, Non-Histone / metabolism
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DNA (Cytosine-5-)-Methyltransferase 1 / genetics
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DNA (Cytosine-5-)-Methyltransferase 1 / metabolism
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DNA (Cytosine-5-)-Methyltransferases / genetics
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DNA (Cytosine-5-)-Methyltransferases / metabolism
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DNA Methyltransferase 3B
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DNA-Binding Proteins / deficiency
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DNA-Binding Proteins / genetics*
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Embryo, Mammalian
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Embryonic Development / genetics*
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Female
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Gene Expression Regulation, Developmental
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Heterochromatin / chemistry
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Heterochromatin / metabolism*
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Histones / genetics
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Histones / metabolism*
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Male
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Methylation
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Methyltransferases / genetics
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Methyltransferases / metabolism
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Mice
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Mice, Knockout
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Mouse Embryonic Stem Cells / cytology
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Mouse Embryonic Stem Cells / metabolism*
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Ovary / abnormalities
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Ovary / growth & development
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Ovary / metabolism
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Primary Cell Culture
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Protein Processing, Post-Translational*
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Testis / abnormalities
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Testis / growth & development
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Testis / metabolism
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X-linked Nuclear Protein / genetics
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X-linked Nuclear Protein / metabolism
Substances
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Chromosomal Proteins, Non-Histone
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DNA-Binding Proteins
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Heterochromatin
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Histones
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RNA, Messenger
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Repressor Proteins
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zinc finger protein 296, mouse
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Chromobox Protein Homolog 5
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Suv39h1 protein, mouse
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Methyltransferases
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DNA (Cytosine-5-)-Methyltransferase 1
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DNA (Cytosine-5-)-Methyltransferases
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Dnmt1 protein, mouse
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Atrx protein, mouse
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X-linked Nuclear Protein