Induction of DNA Methylation by Artificial piRNA Production in Male Germ Cells
| Autor: | Hidetoshi Hasuwa, Chika Ishii, Satomi Kuramochi-Miyagawa, Atsuo Ogura, Daisuke Itou, Toru Nishimura, Toru Nakano, Yoshitaka Fujihara, Yusuke Shiromoto, Narumi Ogonuki, Yukiho Shin-ya |
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| Rok vydání: | 2015 |
| Předmět: |
Male
endocrine system Embryonic Germ Cells Green Fluorescent Proteins Piwi-interacting RNA Biology General Biochemistry Genetics and Molecular Biology Mice Animals Humans RasiRNA RNA Messenger Epigenetics RNA Small Interfering Gene Genetics Regulation of gene expression Agricultural and Biological Sciences(all) Biochemistry Genetics and Molecular Biology(all) urogenital system Gene Expression Profiling Gene Expression Regulation Developmental DNA Methylation Germ Cells DNA demethylation Microscopy Fluorescence DNA methylation General Agricultural and Biological Sciences |
| Zdroj: | Current Biology. 25:901-906 |
| ISSN: | 0960-9822 |
| DOI: | 10.1016/j.cub.2015.01.060 |
| Popis: | Summary Global DNA demethylation and subsequent de novo DNA methylation take place in mammalian male embryonic germ cells [1–3]. P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs), which are germline-specific small RNAs, have been postulated to be critically important for de novo DNA methylation of retrotransposon genes, and many proteins, including PIWI family proteins, play pivotal roles in this process [4–6]. In the embryonic mouse testis, two mouse PIWI proteins, mouse PIWI-like (MILI) and mouse PIWI2 (MIWI2), are involved in the biogenesis of piRNAs through the so-called ping-pong amplification cycle [7–10], and long single-stranded RNAs transcribed from the gene regions of piRNA clusters have been proposed to be the initial material [11–16]. However, it remains unclear whether transcription from the piRNA clusters is required for the biogenesis of piRNAs. To answer this question, we developed a novel artificial piRNA production system by simple expression of sense and antisense EGFP mRNAs in embryonic male germ cells in the piRNA biogenesis phase. EGFP expression was silenced by piRNA-dependent DNA methylation, indicating that concomitant expression of sense and antisense RNA transcripts is necessary and sufficient for piRNA production and subsequent piRNA-dependent gene silencing. In addition, we demonstrated that this artificial piRNA induction paradigm could be applied to an endogenous gene essential for spermatogenesis, DNMT3L [3, 17, 18]. This study not only provides novel insights into the molecular mechanisms of piRNA production, but also presents an innovative strategy for inducing epigenetic modification in germ cells. |
| Databáze: | OpenAIRE |
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