Epigenetic Regulation and small RNAs in Spermatogenesis

• Project Area: The germline: its developmental cycle and epigenome network
• Project/Area Number: 20062007
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: NAKANO Toru 大阪大学, 生命機能研究科, 教授 (00172370)
• Project Period (FY): 2008 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥112,500,000 (Direct Cost: ¥112,500,000); Fiscal Year 2012: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2011: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2010: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2009: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2008: ¥22,500,000 (Direct Cost: ¥22,500,000)
• Keywords: 遺伝子発現制御 / 小分子RNA / 非コードRNA / エピジェネティクス / piRNA / 生殖細胞 / DNAメチル化 / small RNA / 精子形成 / エピジェネティック制御 / 幹細胞 / PIWI / PIWIファミリー / ピストン修飾 / MILI / MIWI / TDRD1

Research Abstract

We analyzed the molecular functions of two mouse PIWI family members, MILI (mouse piwi like) and MIWI2 (mouse piwi 2) and revealed that these two proteins were crucial for the biogenesis of germ cell specific small RNAs, piRNAs (piwi interacting RNA). We also exhibited that MILI and MIWI2 played pivotal roles in de novo DNA methylation of retrotransposon genes in embryonic male germ cells presumably through piRNA. In addition, we showed the molecular functions of MVH (mouse vasa homologue), a germ cell specific RNA helicase and glycerol-3-phosphate acyltransferase 2 (GPAT2) in piRNA-genesis.

Research Products

• [Journal Article] Targeted gene silencing in mouse germ cells by insertion of a homologous DNA into a piRNA generating locus (2013)
  • Author(s): Yamamoto Y,Watanabe T,Hoki Y, Shirane K, Li Y, Ichiiyanagi K, Kuramochi-Miyagawa S, Toyoda A, Fujiyama A, Oginuma M, Suzuki H, Sado T, Nakano T, Sasaki H
  • Journal Title: Genome Res Volume: 23 Issue: 2 Pages: 292-299
  • DOI: 10.1101/gr.137224.112
  • Peer Reviewed
• [Journal Article] GPAT2, a Mitochondrial Outer Membrane Protein in piRNA Biogenesis in Germline Stem Cells (2013)
  • Author(s): Shiromoto Y, Kuramochi-Miyagawa S, Daiba A, Chuma S, Katanaya A, Nishimura K, Ohtaka M, Nakanishi M, Nakamura T, Yoshinaga K, Asada N, Nakamura S, Yasunaga T, Kojima-Kita K, Itou D, Kimura T, Nakano T
  • Journal Title: RNA Volume: 19 Issue: 6 Pages: 803-810
  • DOI: 10.1261/rna.038521.113
  • Peer Reviewed
• [Journal Article] Effects of Dppa3 on DNA methylation dynamics during primordial germ cell development (2013)
  • Author(s): Nakashima H, Kimura T, Kaga Y. Nakatani T, Seki Y, Nakamura T, Nakano T
  • Journal Title: Biol Rep Volume: 88 Issue: 5 Pages: 1-9
  • DOI: 10.1095/biolreprod.112.105932
  • Peer Reviewed
• [Journal Article] GPAT2, a Mitochondrial Outer Membrane Protein, in piRNA Biogenesis in Germline Stem Cells (2013)
  • Author(s): Shiromoto Y, Kuramochi-Miyagawa S, Daiba A, Chuma S, Katanaya A, Nishimura K, Ohtaka M, Nakanishi M, Nakamura T, Yoshinaga K, Asada N, Nakamura S, Yasunaga T, Kojima-Kita K, Itou D, Kimura T, Nakano T
  • Journal Title: RNA Volume: (In press)
  • Peer Reviewed
• [Journal Article] A forward genetic screen to study mammalian RNA interference - essential role of RNase IIIa domain of Dicer1 in 3' strand cleavage of dsRNA in vivo (2012)
  • Author(s): Ohishi K, Nakano T
  • Journal Title: FEBS J. Volume: 279 Issue: 5 Pages: 832-43
  • DOI: 10.1111/j.1742-4658.2012.08474.x
  • Peer Reviewed
• [Journal Article] Essential role of DPPA3 for chromatin condensation in mouse oocytogenesis (2012)
  • Author(s): Liu YJ, Nakamura T, Nakano T
  • Journal Title: Biol Rep Volume: 86 Issue: 2 Pages: 1-8
  • DOI: 10.1095/biolreprod.111.095018
  • Peer Reviewed
• [Journal Article] PGC7 binds histone H3K9me2 to protect against conversion of 5MeC to 5HmC in early embryos (2012)
  • Author(s): Nakamura, T., Liu, Y.U., Nakashima, H., Umehara, H., Inoue, K., Matoba, S., Tachibana, M., Ogura, A., Shinkai, Y., Nakano, T
  • Journal Title: Nature Volume: 486 Issue: 7403 Pages: 415-419
  • DOI: 10.1038/nature11093
  • Peer Reviewed
• [Journal Article] MitoPLD Is a Mitochondrial Protein Essential for Nuage Formation and piRNA Biogenesis in the Mouse Germline (2011)
  • Author(s): Watanabe T, et al
  • Journal Title: Dev Cell Volume: 20 Issue: 3 Pages: 364-375
  • DOI: 10.1016/j.devcel.2011.01.005
  • Peer Reviewed
• [Journal Article] Role for piRNAs and non-coding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus (2011)
  • Author(s): Watanabe, T., Tomizawa, S., Mitsuya, K., Totoki, Y., Yamamoto, Y., Kuramochi-Miyagawa, S., Iida, N., Hoki, Y., Murphy, P.J., Toyoda, A., Gotoh, K., Hiura, H., Arima, T., Fujiyama, A., Sado, T., Shibata, T., Nakano, T., Lin, H., Ichiyanagi, K., Soloway, P.D., Sasaki, H
  • Journal Title: Science Volume: 332 Issue: 6031 Pages: 848-52
  • DOI: 10.1126/science.1203919
  • Peer Reviewed
• [Journal Article] Tudor domain containing 7 (Tdrd7) is essential for dynamic ribonucleoprotein (RNP) remodeling of chromatoid bodies during spermatogenesis (2011)
  • Author(s): Tanaka T, et al
  • Journal Title: Proc Natl Acad Sci USA Volume: 108 Issue: 26 Pages: 10579-10584
  • DOI: 10.1073/pnas.1015447108
  • Peer Reviewed
• [Journal Article] Metastable primordial germ cell-like state induced from mouse embryonic stem cells .by Akt activation (2010)
  • Author(s): Yamano N, Kimura T, Watanabe S, Shinohara T, Nakano T
  • Journal Title: Biochem Biophys Res Commun Volume: 392 Pages: 311-316
  • Peer Reviewed
• [Journal Article] MVH in piRNA processing and gene silencing of retrotransposons (2010)
  • Author(s): Kuramochi-Miyagawa S, Watanabe T, Kimura T, Nakatsuji N, Noce T, Sasaki H, Nakano T, et al
  • Journal Title: Genes Dev Volume: 24 Pages: 887-892
  • Peer Reviewed
• [Journal Article] Disabled-2 is required for mesoderm differentiation of murine embryonic stem cells (2010)
  • Author(s): Huang CL, Cheng JC, Kitajima K, Nakano T, Yeh CF, Chong KY, Tseng CP
  • Journal Title: J Cell Physiol Volume: 225 Pages: 92-105
  • Peer Reviewed
• [Journal Article] MVH in piRNA processing and gene silencing of retrotransposons (2010)
  • Author(s): Kuramochi-Miyagawa S, Nakano T, et al
  • Journal Title: Genes and Development Volume: 24 Pages: 887-892
  • Peer Reviewed
• [Journal Article] Metastable primordial germ cell-like state induced from mouse embryonic stem cells by Akt activation (2010)
  • Author(s): Yamano N, Nakano T, et al
  • Journal Title: Biochem Biophys Res Commun Volume: 392 Pages: 311-316
  • Peer Reviewed
• [Journal Article] MVH in piRNA processing and gene silencing of retrotransposons (2010)
  • Author(s): Kuramochi-Miyagawa S, Watanabe T, Gotoh K, Takamatasu K, Chuma S, Kojima-Kita K, Shiromoto Y, Asada N, Toyoda A, Fujiyama A, Totoki Y, Shibata T, Kimura T, Nakatsuji N, Noce T, Sasaki H, Nakano T.
  • Journal Title: Genes Dev 24 Pages: 887-892
  • Peer Reviewed
• [Journal Article] Associations between PIWI proteins and TDRD1/MTR-1 are critical for integrated subcellular localization in murine male germ cells (2009)
  • Author(s): Kojima K, Nakano T, et al
  • Journal Title: Genes to Cells Volume: 14 Pages: 1155-1165
  • Peer Reviewed
• [Journal Article] Increase of hematopoietic progenitor and suppression of endothelial gene expression by Runx1 expression during in vitro ES differentiation (2009)
  • Author(s): Sakai E, Kitajima K, Sato A, Nakano T
  • Journal Title: Experimental Hematology Volume: 37 Pages: 334-345
  • Peer Reviewed
• [Journal Article] The TDRD9-MIWI2 complex is essential for piRNA-mediated retrotransposon silencing in the mouse male germline (2009)
  • Author(s): Shoji M, Nakano T, et al
  • Journal Title: Developmental Cell Volume: 17 Pages: 775-787
  • Peer Reviewed
• [Journal Article] Gtsf1/Cue110. a gene encoding a protein with two copies of a CHHC Zn-finger motif. is involved in spermatogenesis and retrotransposon suppression in murine testes (2009)
  • Author(s): Yoshimura T, Nakano T, Miyazaki J, et al
  • Journal Title: Developmental Biology Volume: 335 Pages: 216-227
  • Peer Reviewed
• [Journal Article] MILI. a piRNA binding protein. is required for germline stem cell self-renewal and appears to positively regulate translation (2009)
  • Author(s): Unhavaithaya Y, Nakano T, Lin H, et al
  • Journal Title: Journal of Biological Chemistry Volume: 284 Pages: 6507-6519
  • Peer Reviewed
• [Journal Article] MILI, a piRNA binding protein, is required for germline stem cell self-renewal and appears to positively regulate translation. (2009)
  • Author(s): Unhavaithaya Y, Hao Y, Beyret E, Yin H, Kuramochi-Miyagawa S, Nakano T, Lin H.
  • Journal Title: J Biol Chem 284 Pages: 6507-6519
  • Peer Reviewed
• [Journal Article] The TDRD9-MIWI2 complex is essential for piRNA-mediated retrotransposon silencing in the mouse male germline (2009)
  • Author(s): Shoji M, Tanaka T, Hosokawa M, Reuter M, Stark A, Kato Y, Kondoh G, Okawa K, Chujo, Suzuki T, Hata K, Martin S, Noce T, Kuramochi-Miyagawa S, Nakano T, Sasaki H, Pillai RS, Nakatsuji N, Chuma S.
  • Journal Title: Dev Cell 17 Pages: 775-787
  • Peer Reviewed
• [Journal Article] Associations between PIWI proteins and TDRD1/MTR-1 are critical for integrated subcellular localization in murine male germ cells (2009)
  • Author(s): Kojima K, (5名), Nakano T
  • Journal Title: Genes to Cells 14 Pages: 1155-1165
  • Peer Reviewed
• [Journal Article] Increase of hematopoietic progenitor and suppression of endothelial gene expressi on by Runxl expression during in vitro ES differentiation (2009)
  • Author(s): Sakai E, Kitajima K, Sato A, Nakano T
  • Journal Title: Experimental Hematology 37 Pages: 334-345
  • Peer Reviewed
• [Journal Article] The TDRD9-MIWI2 complex is essential for piRNA-mediated retrotransposon silening in the mouse male germline (2009)
  • Author(s): Shoji M, (13名), Nakano T, (4名)
  • Journal Title: Developmental Cell 17 Pages: 775-787
  • Peer Reviewed
• [Journal Article] Gtsf1/Cue110, a gene encoding a protein with two copies of a CHHC Zn-fingermotif, is involved in spermatogenesis and retrotransposon suppression in murine testes (2009)
  • Author(s): Yoshimura T, (6名), Nakano T, Miyazaki J
  • Journal Title: Developmental Biology 335 Pages: 216-227
  • Peer Reviewed
• [Journal Article] MILI, a piRNA binding protein, is required for germline stem cell self-renewal and appears to positively regulate translation (2009)
  • Author(s): Unhavaithaya Y, (4名), Nakano T, Lin H
  • Journal Title: Journal of Biological Chemistry 284 Pages: 6507-6519
  • Peer Reviewed
• [Journal Article] Endogenous siRNAs from naturally formed dsRNAs regulate transcripts in mouse oocytes (2009)
  • Author(s): Watanabe T, Totoki Y, Toyoda A, Kaneda M, Kuramochi-Miyagawa S, Obata Y, Chiba H, Kohara Y, Kono T, Nakano T, Surani MA, Sakaki Y, Sasaki H
  • Journal Title: Nature 453 Pages: 539-543
  • Peer Reviewed
• [Journal Article] Increase of hematopoietic progenitor and suppression of endothelial gene expression by Runxl expression during in vitro ES differentiation (2009)
  • Author(s): Sakai E, Kitajima K, Sato A, Nakano T
  • Journal Title: Exp Hematol 37 Pages: 334-345
  • Peer Reviewed
• [Journal Article] Effects of Akt signaling on nuclear reprogramming (2009)
  • Author(s): Nakamura T, Inoue K, Ogawa S, Umehara H, Ogonuki N, Miki H, Kimura T, Ogura A, Nakano T
  • Journal Title: Genes Cells (in press)
• [Journal Article] MILl, a piRNA binding protein, is required for germline stem cell self-renewal and appears to positively regulate translation (2009)
  • Author(s): Unhavaithaya Y, Hao Y, Beyret E, Yin H, Kuramochi-Miyagawa S, Nakano T, Lin H
  • Journal Title: J Biol Chem (in press)
• [Journal Article] DNA methylation of retrotransposon genes is regulated by Piwi family members MILI and MIWI2 in murine fetal testes. (2008)
  • Author(s): Kuramochi-Miyagawa S, Watanabe T, Gotoh K, Totoki Y, Toyoda A, Ikawa M, Asada N, Kojima K, Yamaguchi Y, Ijiri T, Hata K, Li E, Matsuda Y, Kimura T, Okabe M, Sakaki Y, Sasaki H, Nakano T
  • Journal Title: Genes Dev 22 Pages: 918-930
  • Peer Reviewed
• [Journal Article] DNA methylation of retrotransposon genes is regulated by Piwi family members MILI and MIWI2 in murine fetal testes (2008)
  • Author(s): Kuramochi-Miyagawa S, Watanabe T, Gotoh K, Totoki Y, Toyoda A, Ikawa M, Asada N, Kojima K, Yamaguchi Y, Ijiri T, Hata K, Li E, Matsuda Y, Kimura T, Okabe M, Sakaki Y, Sasaki H, Nakano T
  • Journal Title: Genes Dev 22 Pages: 918-930
  • Peer Reviewed
• [Journal Article] Akt signaling promotes derivation of embryonic germ cells from primordial germ cell (2008)
  • Author(s): Kimura T, Tomooka M, Yamano N, Murayama K, Matoba S, Umehara H, Kanai Y, Nakano T
  • Journal Title: Development 135 Pages: 869-879
  • Peer Reviewed
• [Journal Article] Differential context-dependent effects of FOG-1 on mast cell development and differentiation (2008)
  • Author(s): Sugiyama D, Tanaka M, Kitajima K, Zheng J, Yen H, Murotani T, Yamatodani A, Nakano T
  • Journal Title: Blood 111 Pages: 1924-1932
  • Peer Reviewed
• [Journal Article] Calyculin A retraction of mature megakaryocytes proplatelets from embryonic stem cells (2008)
  • Author(s): Tamaru S, Kitajima K, Nakano T, Eto K, Yazaki A, Kobayashi T, Matsuinoto T, Wada H, Katayama N, Nishikawa M
  • Journal Title: Biochem Biophys Res Commun 366 Pages: 763-768
  • Peer Reviewed
• [Journal Article] PI3K/Akt signaling as a key regulatory pathway for chondrocyte terminal differentiation (2008)
  • Author(s): Kita K, Kimura T, Nakamura N, Yoshikawa H, Nakano T
  • Journal Title: Genes Cells 13 Pages: 839-850
  • Peer Reviewed
• [Journal Article] MVH in piRNA processing and gene silencing of retrotransposons
  • Author(s): Kuramochi-Miyagawa S, (15名), Nakano T
  • Journal Title: Genes and Development (In press)
  • Peer Reviewed
• [Journal Article] Metastable primordial germ cell-like state induced from mouse embryonic stem cells by Akt activation
  • Author(s): Yamano N, (3名), Nakano T
  • Journal Title: Biochem Biophys Res Commun (In press)
  • Peer Reviewed
• [Presentation] Artilicial Incluction of Germ Cell Specific liene Sileucing through piRNA pathway (2013)
  • Author(s): 仲野 徹
  • Organizer: Small RNAs to Stem Cells & Epigenetic Reprogramming Asia-2013
  • Place of Presentation: 東京大学、東京
  • Year and Date: 2013-11-25
  • Invited
• [Presentation] Artificial Induction of Germ Cell Specific Gene Silencing tnrough piRNA pathway (2013)
  • Author(s): 仲野 徹
  • Organizer: 8th Annual Conference of Asia Epigenome Alliance
  • Place of Presentation: 台北(台湾)
  • Year and Date: 2013-11-06
  • Invited
• [Presentation] Artificial Induction of Germ cell Specific Gene Silencing tnrouga piRNA pathway (2013)
  • Author(s): 仲野 徹
  • Organizer: Recent Advances in Stem Cell Biology
  • Place of Presentation: 九州大学、博多
  • Year and Date: 2013-11-04
  • Invited
• [Presentation] DMメチル化と発生・発がん (2013)
  • Author(s): 仲野 徹
  • Organizer: 日本病理学会
  • Place of Presentation: 札幌
  • Year and Date: 2013-06-07
  • Invited
• [Presentation] piRNA産生システムの解析 (2012)
  • Author(s): 仲野 徹
  • Organizer: 22nd HCS 4th JARI Joint International Symposium
  • Place of Presentation: 広島市
  • Year and Date: 2012-08-30
• [Presentation] piRNA産生システムの解析 (2012)
  • Author(s): 仲野徹
  • Organizer: 22nd HCS 4th JARI Joint International Symposium
  • Place of Presentation: 広島
  • Year and Date: 2012-08-30
• [Presentation] piRNA and Spermatogenesis (2012)
  • Author(s): 仲野 徹
  • Organizer: 2nd SKLRB Symposia
  • Place of Presentation: 北京・中国
  • Year and Date: 2012-05-07
• [Presentation] DNA Methylation in Cell Differentiation and Transformation (2012)
  • Author(s): 仲野 徹
  • Organizer: Shanghai International Conference of Epigenetics in Development and Disease
  • Place of Presentation: 上海・中国
  • Year and Date: 2012-04-21
• [Presentation] piRNA産生システムの解析 (2011)
  • Author(s): 仲野 徹
  • Organizer: 特定領域研究 生殖サイクル 公開シンポジウム
  • Place of Presentation: 大阪市
  • Year and Date: 2011-11-17
• [Presentation] DNA methylation control by PGC7/Stella (2010)
  • Author(s): 仲野 徹
  • Organizer: BMB 2010
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-07
• [Presentation] DNA methylation control by PGC7/Stella (2010)
  • Author(s): 仲野徹
  • Organizer: BMB 2010
  • Place of Presentation: 神戸(招待講演)
  • Year and Date: 2010-12-07
• [Presentation] 細胞分化・がん化におけるエピジェネティクス制御 (2010)
  • Author(s): 仲野 徹
  • Organizer: JDDW2010
  • Place of Presentation: 横浜市
  • Year and Date: 2010-10-15
• [Presentation] 細胞分化・がん化におけるエピジェネティクス制御 (2010)
  • Author(s): 仲野徹
  • Organizer: JDDW2010
  • Place of Presentation: 横浜(招待講演)
  • Year and Date: 2010-10-15
• [Presentation] 幹細胞の未分化性とシグナル (2010)
  • Author(s): 仲野 徹
  • Organizer: 第69回日本癌学会
  • Place of Presentation: 大阪市
  • Year and Date: 2010-09-23
• [Presentation] 幹細胞の未分化性とシグナル (2010)
  • Author(s): 仲野徹
  • Organizer: 第69回日本癌学会
  • Place of Presentation: 大阪(招待講演)
  • Year and Date: 2010-09-23
• [Presentation] DNA methylation in Early Embryogenesis (2010)
  • Author(s): 仲野 徹
  • Organizer: Epigenetics : New horizons in Japan and Scandinavia
  • Place of Presentation: ストックホルム・スウェーデン
  • Year and Date: 2010-09-06
• [Presentation] DNA methylation in Early Embryogenesis (2010)
  • Author(s): 仲野徹
  • Organizer: Epigenetics : New horizons in Japan and Scandinavia
  • Place of Presentation: ストックホルム スウェーデン(招待講演)
  • Year and Date: 2010-09-06
• [Presentation] 発生・分化におけるエピジェネティクス制御 (2009)
  • Author(s): 仲野 徹
  • Organizer: 日本免疫学会
  • Place of Presentation: 大阪市
  • Year and Date: 2009-12-03
• [Presentation] Epigenetic gene regulation and GATA factors (2009)
  • Author(s): 仲野 徹
  • Organizer: International Society of Experimental Hematology
  • Place of Presentation: アテネ・ギリシャ
  • Year and Date: 2009-09-11
• [Presentation] Epigenetic gene regulation and GATA factors (2009)
  • Author(s): 仲野徹
  • Organizer: International Society of Experimental Hematology
  • Place of Presentation: アテネ、ギリシャ
  • Year and Date: 2009-09-11
• [Presentation] DNA methylation in early embryogenesis (2009)
  • Author(s): 仲野 徹
  • Organizer: From Imprinting to the Epigenome
  • Place of Presentation: ケンブリッジ・イギリス
  • Year and Date: 2009-09-04
• [Presentation] DNA methylation in early embryogenesis (2009)
  • Author(s): 仲野徹
  • Organizer: From Imprinting to the Epigenome
  • Place of Presentation: ケンブリッジ、英国
  • Year and Date: 2009-09-04
• [Presentation] DNA methylation in early embryogenesis (2009)
  • Author(s): 仲野 徹
  • Organizer: 日本発生生物学会
  • Place of Presentation: 新潟市
  • Year and Date: 2009-05-29
• [Presentation] Small RNA and Gene Silencing in Spermatogenesis (2009)
  • Author(s): 仲野徹
  • Organizer: 9th International Congress of Andrology
  • Place of Presentation: バルセロナ、スペイン
  • Year and Date: 2009-03-09
• [Presentation] DNA methylation in early embryogenesis (2008)
  • Author(s): 仲野徹
  • Organizer: Decoding Epigenetic Code
  • Place of Presentation: 東京
  • Year and Date: 2008-12-15
• [Presentation] Regulation of Stem Cell Systems by PI3K/Akt Signaling (2008)
  • Author(s): Nakano T
  • Organizer: The 5th Nikko International Symposium : Stem Cells and Cancer
  • Place of Presentation: Tochigi
  • Year and Date: 2008-10-25
• [Presentation] DNA methylation in early embryogenesis (2008)
  • Author(s): 仲野徹
  • Organizer: Royan International Twin Congress
  • Place of Presentation: テヘラン、イラン
  • Year and Date: 2008-08-28
• [Presentation] DNAメチル化制御とsmall RNA (2008)
  • Author(s): 仲野徹
  • Organizer: 第二回エピジェネティクス研究会
  • Place of Presentation: 三島、静岡県
  • Year and Date: 2008-05-09
• [Remarks]
  • URL: http://www.fbs.osaka-u.ac.jp/labs/nakano/researches.html