Zfp296 negatively regulates H3K9 methylation in embryonic development as a component of heterochromatin

• Published in: Scientific reports Vol. 7; no. 1; pp. 12462 - 12
• Main Authors: Matsuura, Takumi, Miyazaki, Satsuki, Miyazaki, Tatsushi, Tashiro, Fumi, Miyazaki, Jun-ichi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 29.09.2017; Nature Publishing Group
• Subjects: 13/51; 42/41; 45/77; 631/136/1455; 631/337/176/2016; 64/60; 82/58; 82/80; 96/1; Ablation; Animals; Cell Differentiation; Chromobox Protein Homolog 5; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; Developmental stages; DNA (Cytosine-5-)-Methyltransferase 1 - genetics; DNA (Cytosine-5-)-Methyltransferase 1 - metabolism; DNA (Cytosine-5-)-Methyltransferases - genetics; DNA (Cytosine-5-)-Methyltransferases - metabolism; DNA Methyltransferase 3B; DNA-Binding Proteins - deficiency; DNA-Binding Proteins - genetics; DNMT1 protein; Embryo, Mammalian; Embryogenesis; Embryonic Development - genetics; Embryonic growth stage; Embryos; Female; Fertility; Gene Expression Regulation, Developmental; Heterochromatin; Heterochromatin - chemistry; Heterochromatin - metabolism; Histone deacetylase; Histones - genetics; Histones - metabolism; Humanities and Social Sciences; Leukemia; Male; Methylation; Methyltransferases - genetics; Methyltransferases - metabolism; Mice; Mice, Knockout; Mouse Embryonic Stem Cells - cytology; Mouse Embryonic Stem Cells - metabolism; multidisciplinary; NuRD protein; Ovaries; Ovary - abnormalities; Ovary - growth & development; Ovary - metabolism; Pluripotency; Primary Cell Culture; Protein Processing, Post-Translational; Proteins; Repressor Proteins - genetics; Repressor Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Science; Science (multidisciplinary); Sperm; Stem cells; Testis - abnormalities; Testis - growth & development; Testis - metabolism; Tumorigenesis; X-linked Nuclear Protein - genetics; X-linked Nuclear Protein - metabolism; Zinc finger proteins
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-12772-y

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.