piRNA biogenesis pathway in mouse spermatogenesis

• Project/Area Number: 24370074
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Osaka University
• Principal Investigator: Miyagawa Satomi 大阪大学, 医学(系)研究科(研究院), 特任講師(常勤) (90291153)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2014: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: piRNA / 小分子RNA / 精子形成 / レトロトランスポゾン / ミトコンドリア / 生殖顆粒 / 発生・分化 / PIWI / タンパク質 / 細胞・組織

Outline of Final Research Achievements

piRNA (PIWI interacting RNA) is a germ cell-specific small RNA in whose biogenesis PIWI (P element wimpy testis) family proteins play crucial roles. MILI (mouse Piwi like), one of the mouse PIWI family members, is indispensable for the piRNA production, DNA methylation of retrotransposons through the piRNA, and spermatogenesis. To analyze the molecular function of MILI in piRNA biogenesis, we utilized germline stem cells (GS cells), which are derived from testicular stem cells and possess spermatogonial phenotype. We found that glycerol-3-phosphate acyltransferase 2 (GPAT2), a mitochondrial outer membrane protein for lysophosphatidic acid, bound to MILI and that gene knockdown of GPAT2 brought about the impaired piRNA production in GS cells. GPAT2 is not only one of the MILI bound proteins but also a protein essential for the primary piRNA biogenesis. In addition, MVH (mouse vasa homologues) helicase activity is also indispensable for piRNA production and spermatogenesis.

Research Products

• [Journal Article] Reply to Shoji and Katsuma (2015)
  • Author(s): Kuramochi-Miyagawa S, Nakano T
  • Journal Title: Current Biology Volume: 25 Pages: 901-906
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive DNA Methylation Analysis of Retrotransposons in Male Germ Cells (2015)
  • Author(s): Nagamori I, Kobayashi H, Shiromoto Y, Nishimura T, Kuramochi-Miyagawa S, Kono T, Nakano T.
  • Journal Title: Cell Reports Volume: 12 Pages: 1541-1547
  • DOI: 10.1016/j.celrep.2015.07.060
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] piRNAs derived from ancient viral processed pseudogenes as transgenerational sequence-specific immune memory in mammals. (2015)
  • Author(s): Parrish NF, Fujino K, Shiromoto Y, Iwasaki YW, Ha H, Xing J, Makino A, Kuramochi-Miyagawa S, Nakano T, Siomi H, Honda T, Tomonaga K.
  • Journal Title: RNA Volume: 21 Pages: 1691-1703
  • DOI: 10.1261/rna.052092.115
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Induction of DNA Methylation by Artificial piRNA Production in Male Germ Cells (2015)
  • Author(s): Itou D, Shiromoto Y, Shin-Ya Y, Ishii C, Nishimura T, Ogonuki N, Ogura A, Hasuwa H, Fujihara Y, Kuramochi-Miyagawa S, Nakano T.
  • Journal Title: Current Biol Volume: 25 Pages: 901-6
  • DOI: 10.1016/j.cub.2015.01.060
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] HSP90α plays an important role in piRNA biogenesis and retrotransposon repression in mouse (2014)
  • Author(s): Ichiyanagi, T., Ichiyanagi, K., Ogawa, A., Kuramochi-Miyagawa, S., Nakano, T., Chuma, S., Sasaki, H.
  • Journal Title: Nucleic Acids Res. Volume: 42 Pages: 11903-11
  • DOI: 10.1093/nar/gku881
  • NAID: 120005496571
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Analysis of DNA methylation in Mouse Testis (2014)
  • Author(s): Kuramochi-Miyagawa S, Kojima-Kita K, Shiromoto Y, Itou D, Koshima H, Nakano T
  • Journal Title: Methods Mol Biol. Volume: 1093 Pages: 97-109
  • DOI: 10.1007/978-1-62703-694-8_8
  • Peer Reviewed / Open Access
• [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 Pages: 803-10
  • DOI: 10.1261/rna.038521.113
  • Peer Reviewed
• [Journal Article] The nuage mediates retrotransposon silencing in mouse primordial ovarian follicles (2013)
  • Author(s): Lim AK, Lorthongpanich C, Chew TG, Tan CW, Shue YT, Balu S, Gounko N, Kuramochi-Miyagawa S, Matzuk MM, Chuma S, Messerschmidt DM, Solter D, Knowles BB
  • Journal Title: Development. Volume: 140(18) Pages: 3819-25
  • DOI: 10.1242/dev.099184
  • Peer Reviewed
• [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 Pages: 292-299
  • DOI: 10.1101/gr.137224.112
  • Peer Reviewed
• [Journal Article] Identification of MIWI-associated Poly(A) RNAs by immunoprecipitation with an anti-MIWI monoclonal antibody (2012)
  • Author(s): Nishibu, T., Hayashida, Y., Tani, S., Kurono, S., Kojima-Kita, K., Ukekawa, R., Kurokawa, T., Kuramochi-Miyagawa, S., Nakano, T., Inoue, K., and Honda, S.
  • Journal Title: BST Volume: 6 Issue: 5 Pages: 248-261
  • DOI: 10.5582/bst.2012.v6.5.248
  • NAID: 130005054634
  • ISSN: 1881-7815, 1881-7823
  • Peer Reviewed
• [Presentation] 雄性生殖細胞における人為的piRNAを介したDNAメチル化の誘導 (2015)
  • Author(s): 伊藤大介、新屋幸穂、城本悠助,、宮川さとみ、仲野徹
  • Organizer: 第９回日本エピジェネティクス研究会
  • Place of Presentation: 学術総合センター(東京都）
  • Year and Date: 2015-05-25
• [Presentation] 雄性生殖細胞における人為的piRNAを介したDNAメチル化の誘導 (2014)
  • Author(s): 伊藤大介、城本悠助、宮川さとみ、仲野徹
  • Organizer: 第16回日本RNA学会年会
  • Place of Presentation: ウィンク愛知（愛知県名古屋市）
  • Year and Date: 2014-07-23
• [Presentation] 雄性生殖細胞における人為的piRNAを介したDNAメチル化の誘導 (2014)
  • Author(s): 伊藤大介、城本悠助、宮川さとみ、仲野徹
  • Organizer: 生殖エピゲノム若手勉強会2014
  • Place of Presentation: つくばグランドホテル(茨城県つくば市)
  • Year and Date: 2014-07-16
• [Presentation] ミトコンドリア外膜タンパクGPAT2のpiRNA産生における機能 (2013)
  • Author(s): 城本悠助、宮川（倉持）さとみ、仲野徹
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2013-12-03 – 2013-12-06
• [Presentation] MVHヘリカーゼ活性はLINE-1レトロトランスポゾンの抑制に必須である (2013)
  • Author(s): 伊藤大介、宮川（倉持）さとみ、仲野徹、
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2013-12-03 – 2013-12-06
• [Presentation] 雄性生殖細胞におけるpiRNA依存的遺伝子サイレンシングの人為的誘導 (2013)
  • Author(s): 新屋幸穂、伊藤大介、宮川（倉持）さとみ、仲野徹
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2013-12-03 – 2013-12-06
• [Presentation] 新学術領域『非コードRNA作用マシナリー』領域班会議 (2013)
  • Author(s): 伊藤大介、宮川（倉持）さとみ、城本悠助、仲野徹
  • Organizer: 第7回日本エピジェネティクス研究会年会
  • Place of Presentation: 奈良
  • Year and Date: 2013-05-30 – 2013-05-31
• [Presentation] マウス精巣におけるpiRNAを介した配列特異的なDNAメチル化誘導システム
  • Author(s): 伊藤大介、宮川さとみ、城本悠助、仲野徹
  • Organizer: 第6回 日本エピジェネティクス研究会年会
  • Place of Presentation: 東京都 一ツ橋学術総合センター
• [Presentation] 精子幹細胞を用いたpiRNA産生機構の解析
  • Author(s): 城本悠助、宮川さとみ、他6名
  • Organizer: NGS現場の会 第二回研究会
  • Place of Presentation: 大阪府吹田市 ホテル阪急エキスポパーク
• [Presentation] 精子幹細胞を用いたpiRNA産生機構の解析
  • Author(s): 城本悠助、宮川さとみ、他4名
  • Organizer: 特定領域生殖サイクル若手勉強会 2012
  • Place of Presentation: 宮城県仙台市 秋保リゾート ホテルクレセント
• [Presentation] GPAT2, a mitochondrial acyltransferase, in piRNA biogenesis in germline stem cells.
  • Author(s): Yusuke Shiromoto, et.el
  • Organizer: REGULATORY & NON-CODING RNAs in Cold Spring Habor Laboratory meeting
  • Place of Presentation: Cold Spring Habor Laboratory, New York, USA
• [Presentation] De novo DNA methylation by artificially produced piRNA in murine teste
  • Author(s): Itou D, et.el
  • Organizer: Epigenetics and Chromatin in Cold Spring Habor Laboratory meeting
  • Place of Presentation: Cold Spring Habor Laboratory, New York, USA
• [Presentation] マウス精巣における人為的piRNA産生を介したDNAメチル化制御
  • Author(s): 伊藤大介、宮川さとみ、城本悠助、仲野徹
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡市 福岡国際会議場・マリンメッセ福岡