配偶子ゲノムの修飾機構と胚発生への影響

• Project/Area Number: 15080208
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute of Genetics
• Principal Investigator: 佐々木 裕之 National Institute of Genetics, 総合遺伝研究系, 教授 (30183825)
• Co-Investigator(Kenkyū-buntansha): 佐渡 敬 国立遺伝学研究所, 総合遺伝研究系, 助教 (70321601) ; 秦 健一郎 国立育成医療センター(研究所), 周産期病態研究部, 部長 (60360335)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥120,600,000 (Direct Cost: ¥120,600,000); Fiscal Year 2007: ¥21,900,000 (Direct Cost: ¥21,900,000); Fiscal Year 2006: ¥22,200,000 (Direct Cost: ¥22,200,000); Fiscal Year 2005: ¥25,500,000 (Direct Cost: ¥25,500,000); Fiscal Year 2004: ¥25,500,000 (Direct Cost: ¥25,500,000); Fiscal Year 2003: ¥25,500,000 (Direct Cost: ¥25,500,000)
• Keywords: 発生・分化 / 遺伝学 / ゲノムインプリンティング / DNAメチル化

Research Abstract

最終年度にあたり、研究計画にそって以下の研究を行い、結果を取りまとめた。(1)DNAメチル化酵素Dnmt3aと補助因子Dnmt3Lに加えて、Dnmt3bが雄の配偶子形成過程で反復配列を修飾することが分かったので、その詳細な解析を完成させ報告した。また、生殖細胞特異的に発現するpiRNAに結合するPiwiファミリー蛋白質が、反復配列(とくにレトロトランスポソン)のメチル化に関わることを発見し報告した。(2)一方、Dnmt3bの欠損は雌雄の配偶子形成を阻害せず、仔の発生にも影響がないことが分かった。すなわち、反復配列のメチル化は受精後のDnmt3bの供給により回復される。現在投稿準備中である。(3)ゲノムワイドな脱メチル化が生じる卵割期に(Dnmt3aやDnmt3bではなく)Dnmt1がインプリントを維持することを証明できたので、論文にまとめた(投稿中)。最後に、(4)two-hybrid法にて上記メチル化酵素複合体と相互作用する因子を得るため、生殖細胞特異的ライブラリーを作成した。このライブラリーは、将来の研究において役立てる予定である。

Research Products

• [Journal Article] Epigenetic events in mammalian germ-cell development: reprog ramming and beyond (2008)
  • Author(s): Sasaki, H. and Matsui Y.
  • Journal Title: Nat Rev. Genet. 9 Pages: 129-140
  • 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., et. al.
  • Journal Title: Gens Dev. 22 Pages: 908-917
  • Peer Reviewed
• [Journal Article] Endogenous siRNAs from naturally formed dsRNAs regulate transcripts in mouse oocytes (2008)
  • Author(s): Watanabe, T., et. al.
  • Journal Title: Nature (In press)
  • Peer Reviewed
• [Journal Article] Identification of the imprinted KLF14 transcription factor undergoing human-specific accelerated evolution (2007)
  • Author(s): Parker-Katiraee, L., et. al.
  • Journal Title: PLoS Genet. 3 Pages: 665-678
  • Peer Reviewed
• [Journal Article] Role of the Dnmt3 family in de novo methylation of imprinted and repetitive sequences during male germ cell development in the mouse (2007)
  • Author(s): Kato, Y., et. al.
  • Journal Title: Hum. Mol. Genet. 16 Pages: 2272-2280
  • Peer Reviewed
• [Journal Article] Analysis of small RNA profiles during development (2007)
  • Author(s): Watanabe, T., et. al.
  • Journal Title: Methods Enzymol. 427 Pages: 155-169
• [Journal Article] Aberrant DNA methylation of imprinted loci in sperm from oligospermic patients (2007)
  • Author(s): Kobayashi, H., et. al.
  • Journal Title: Hum. Mol. Genet. 16 Pages: 2542-2551
  • Peer Reviewed
• [Journal Article] Evolution of the vertebrate DNMT3 gene family : a possible link between existence of DNMT3L and genomic imprinting. (2006)
  • Author(s): Yokomine, T. et al.
  • Journal Title: Cytogenet. Genome Res. 113 Pages: 75-80
• [Journal Article] Bisulfite sequencing and dinucleotide content analysis of 15 imprinted mouse dififerentially methylated regions (DMRs) : paternally methylated DMRs contain less CpGs than maternally methylated DMRs. (2006)
  • Author(s): Kobayashi, H. et al.
  • Journal Title: Cytogenet. Genome Res. 113 Pages: 130-137
• [Journal Article] Loss of the maternal imprint in Dnmt3L^<mat-/-> mice leads to a differentiation defect in the extraembryonic tissue. (2006)
  • Author(s): Arima, T. et al.
  • Journal Title: Dev. Biol. 297 Pages: 361-373
• [Journal Article] Identification and characterization of two novel classes of small RNAs in the mouse germline : retrotransposon-derived siRNAs in oocytes and germline small RNAs in testes. (2006)
  • Author(s): Watanabe, T. et al.
  • Journal Title: Genes Dev. 20 Pages: 1732-1743
• [Journal Article] Meiotic and epigenetic aberrations in Dnmt3L-deficient male germ cells. (2006)
  • Author(s): Hata, K.et al.
  • Journal Title: Mol.Reprod.Dev. 73 Pages: 116-122
• [Journal Article] Stochastic imprinting in the progeny of Dnmt3L-/- females. (2006)
  • Author(s): Arnaud, P.et al.
  • Journal Title: Hum.Mol.Genet. 15 Pages: 589-598
• [Journal Article] Bisulfite sequencing and dinucleotide content analysis of 15 imprinted mouse differentially methylated regions (DMRs) : paternally methylated DMRs contain less CpGs than maternally methylated DMRs. (2006)
  • Author(s): Kobayashi, H.et al.
  • Journal Title: Cytogenet.Genome Res. (in press)
• [Journal Article] Evolution of the vertebrate DNMT3 gene family : a possible link between existence of DNMT3L and genomic imprinting. (2006)
  • Author(s): Yokomine, T.et al.
  • Journal Title: Cytogenet.Genome Res. (in press)
• [Journal Article] Structural and functional analysis of a 0.5-Mb chicken region orthologous to the imprinted mammalian Ascl2/Mash2-Igf2-H19 region (2005)
  • Author(s): Yokomine, T., et al.
  • Journal Title: Genome Res. 15 Pages: 154-165
• [Journal Article] The two domain hypothesis in Beckwith-Wiedemann syndrome : autonomous imprinting of the telomeric domain of the distal chromosome 7 cluster (2005)
  • Author(s): Cerrato, F., et al.
  • Journal Title: Hum.Mol.Genet. 14 Pages: 503-511
• [Journal Article] Imprinting and looping : epigenetic marks control interactions between regulatory elements (2005)
  • Author(s): Kato, Y., et al.
  • Journal Title: BioEssays 27 Pages: 1-5
• [Journal Article] Large-scale identification and mapping of nuclear matrix-attachment regions in the distal imprinted domain of mouse chromosome 7 (2004)
  • Author(s): Purbowasito, W., et al.
  • Journal Title: DNA Res. 11 Pages: 391-407
• [Journal Article] A 210-kb segment of tandem repeats and retroelements located between imprinted subdomains of mouse distal chromosome 7 (2004)
  • Author(s): Shirohzu, H., et al.
  • Journal Title: DNA Res. 11 Pages: 325-334
• [Journal Article] X-inactivation is stably maintained in mouse embryos deficient for histone methyltransferase G9a (2004)
  • Author(s): Ohata, T., et al.
  • Journal Title: Genesis 40 Pages: 151-156
• [Journal Article] Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting (2004)
  • Author(s): Kaneda, M., et al.
  • Journal Title: Nature 429 Pages: 900-903
• [Journal Article] Identification of the imprinted KLF14 transcription factor undergoing human-specific accelerated evolution.
  • Author(s): Parker-Katiraee, L. et al.
  • Journal Title: PLoS Genet. (in press)
• [Journal Article] Role of de novo DNA methyltransferases in initiation of genomic imprinting and X-chromosome inactivation
  • Author(s): Keneda, M., et al.
  • Journal Title: Cold Spring Harbor Symp.Quant.Biol. (in press)
• [Presentation] Endogenous siRNAs derived from naturally formed dsRNAs regulate coding-transcripts and retrotransposons in mouse oocytes (2008)
  • Author(s): Sasaki, H.
  • Organizer: NIBB-EMBL Joint Meeting on Evolution of Epigenetic Regulation
  • Place of Presentation: Heidelberg, Germany
• [Presentation] Epigenomics in reproduction and development (2007)
  • Author(s): Sasaki, H.
  • Organizer: The Asian Network of Epigenomics
  • Place of Presentation: Suita
  • Year and Date: 2007-06-14
• [Presentation] エピジェネティクスと発生 (2007)
  • Author(s): 佐々木 裕之
  • Organizer: 第143回 日本獣医学会大会
  • Place of Presentation: つくば
• [Presentation] Genome imprinting mechanisms in the mouse germline and early embryos (2007)
  • Author(s): Sasaki, H.
  • Organizer: Gordon Research Conference: Epigenetics
  • Place of Presentation: New Hampshire, USA
• [Presentation] 流産のエピゲノム解析 (2007)
  • Author(s): 秦健一郎, ほか
  • Organizer: 第52回人類遺伝学会
  • Place of Presentation: 東京
• [Presentation] Role of DNA methyltransferases(Dnmts)in maintenance of the methylation imprint during mouse pre-implantation development (2007)
  • Author(s): Hirasawa, R., et. al.
  • Organizer: Wellcome Trust Conference on Mouse Molecular Genetics
  • Place of Presentation: Hinxton, UK
• [Presentation] 哺乳類の生殖細胞におけるゲノムインプリンティング機構 (2007)
  • Author(s): 佐々木裕之
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] 男性不妊症の精子におけるインプリント異常 (2007)
  • Author(s): 小林久人, ほか
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] マウス卵における内在性siRNAの生合成経路と機能 (2007)
  • Author(s): 渡部聡朗, ほか
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] マウスPIWIファミリーとエピジェネティクス制御 (2007)
  • Author(s): 宮川さとみ, ほか
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] マウス着床前胚におけるゲノムインプリントの維持に関わるDNAメチル化酵素の解析 (2007)
  • Author(s): 平澤竜太郎, ほか
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] DNA methyltransferases involved in establishment of maternal imprint in mouse oocytes (2007)
  • Author(s): 千葉初音, ほか
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Book] Genomic Imprinting (Special Issue of Cytogenet. Genome Res.) (2006)
  • Author(s): Sasaki, H., Ishino, F.et al.
  • Total Pages: 350
  • Publisher: S. Kargar AG, Basel
• [Book] Genomic Imprinting (Special Issue of Cytogenet.Genome Res.) (2006)
  • Author(s): Sasaki, H., Ishino, F.
  • Publisher: S.Kargar AG, Basel
• [Book] 再生医療教科書シリーズ,第3巻再生医療のための分子生物学(分担) (2006)
  • Author(s): 平澤竜太郎, 佐々木裕之
  • Total Pages: 17
  • Publisher: コロナ社
• [Book] 図説ARTマニュアル(改訂第2版)(分担) (2006)
  • Author(s): 久須美真紀, 佐々木裕之
  • Publisher: 永井書店(印刷中)
• [Book] Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics(分担) (2005)
  • Author(s): Sasaki, H.
  • Publisher: John Wiley & Sons, Chichester
• [Book] エピジェネティクス入門-三毛猫の模様はどう決まるのか (2005)
  • Author(s): 佐々木裕之
  • Total Pages: 92
  • Publisher: 岩波書店
• [Book] エピジェネティクス (2004)
  • Author(s): 佐々木裕之 編
  • Total Pages: 238
  • Publisher: シュプリンガー・フェアラーク東京
• [Book] Analysis of small RNA profiles during development.
  • Author(s): Watanabe, T. et al.
  • Publisher: Methods Enzymol. (in press)
• [Book] Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics (eds.Dunn, M., Jorde, L., Little, P.& Subramaniam, S.)
  • Author(s): Sasaki, H.(分担)
  • Publisher: John Wiley & Sons(印刷中)
• [Publications] Hata, K.: "Genomic imprinting : mechanisms, significance and evolution."J.Mamm.Ova Res.. 20. 64-68 (2003)