Epigenetic Regulation in Development and Differentiation

• Project/Area Number: 21249016
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• 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): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥47,970,000 (Direct Cost: ¥36,900,000、Indirect Cost: ¥11,070,000); Fiscal Year 2011: ¥14,820,000 (Direct Cost: ¥11,400,000、Indirect Cost: ¥3,420,000); Fiscal Year 2010: ¥15,730,000 (Direct Cost: ¥12,100,000、Indirect Cost: ¥3,630,000); Fiscal Year 2009: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
• Keywords: DNAメチル化 / ヒストン修飾 / エピジェネティクス / クロマチン / 遺伝子発現 / 悪性腫瘍 / 初期発生 / メチル化シトシン / ハイドロキシメチル化シトシン / 細胞分化 / 小分子RNA / 精子形成

Research Abstract

Establishment of epigenetic state is critically important for development and differentiation of multicellular organisms. In this project, we carried out following three issues on the establishment of epigenetic status.
1. Function of PGC7/ Stella, a regulator of DNA methylation in early embryos.: PGC7/ Stella has the effect to prevent" active DNA demethylation" in early embryos. We identified that the binding of PGC/ Stella to H3K9me2(dimethylated lysine 9 of histone H3) is critically important for the function. During the course of this project, Tet protein plays critical roles in active DNA demethylation by converting 5-methyl cytosine to 5-hydroxymethyl cytosine. Based on this data, we revealed that the binding of Tet-3, the most abundant Tet protein in early embryos, to chromatin is inhibited by PGC7.
2. We analyzed the function of GATA-1 transcription factor using in vitro hematopoietic differentiation induction system from ES cells. Controlled expression of GATA-1 at different time points showed that the function of the factor is quite varied dependent on the time point and its different roles were controlled by epigenetic mechanisms.
3. We find that GS cells(Germline stem cells) are a quite useful resource for studying the biogenesis of piRNA and that the initial phase of piRNA production is carried out in the cells. Using GS cells, we succeeded to identify a protein involved in piRNA biogenesis. In addition, we established a novel experimental system to induce piRNA and subsequent DNA methylation in the transgenic mice.

Research Products

• [Journal Article] PGC7 binds histone H3K9me2 to protect against conversion of 5MeC to 5HmC in early embryos (2012)
  • Author(s): Nakamura T, Liu YU, Nakashima H, UmeharaH, Inoue K, Matoba S, Tachibana M, OguraA, Shinkai Y, Nakano T.
  • Journal Title: Nature Volume: (in press)
  • Peer Reviewed
• [Journal Article] Essential role of DPPA3 for chromatin condensation in mouse oocytogenesis (2012)
  • Author(s): Ohishi K, Nakano T.
  • Journal Title: FEBS J Volume: 279 Pages: 832-43
  • 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 Pages: 1-8
  • 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: (in press)
  • Peer Reviewed
• [Journal Article] PGC7/Stella links H3K9me2 to protect against conversion of 5MeC to 5HmC in early embryos (2012)
  • Author(s): Nakamura T, et al
  • Journal Title: Nature Volume: (in press)
  • Peer Reviewed
• [Journal Article] RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos. (2011)
  • Author(s): Matoba S, Inoue K, Kohda T, Sugimoto M, Mizutani E, Ogonuki N, Nakamura T, Abe K, Nakano T, Ishino F, Ogura A
  • Journal Title: Proc Natl Acad Sci USA Volume: 108 Pages: 20621-6
  • Peer Reviewed
• [Journal Article] Locus-and domain-dependent control of DNA methylation at mouse B1 retrotransposons during male germ cell development (2011)
  • Author(s): Ichiyanagi K, Li Y, Watanabe T, Ichiyanagi T, Fukuda K, Kitayama J, Yamamoto Y, Kuramochi-Miyagawa S, NakanoT, Yabuta Y, Seki Y, Saitou M, Sasaki H
  • Journal Title: Genome Res Volume: 21 Pages: 2058-66
  • 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, Hosokawa M, Vagin VV, ReuterM, Hayashi E, Mochizuki AL, Kitamura K, Yamanaka H, Kondoh G, Okawa K, Kuramochi-Miyagawa S, Nakano T, Sachidanandam R, Hannon GJ, Pillai RS, Nakatsuji N, Chuma S
  • Journal Title: Proc Natl Acad Sci USA Volume: 108 Pages: 10579-10584
  • Peer Reviewed
• [Journal Article] Regulation of the MDM2-P53 pathway and tumor growth by PICT1 via nucleolar RPL11 (2011)
  • Author(s): Sasaki M, Kawahara K, Nishio M, MimoriK, Kogo R, Hamada K, Itoh B, Wang J, KomatsuY, Yang YR, Hikasa H, Horie Y, YamashitaT, Kamijo T, Zhang Y, Zhu Y, Prives C, Nakano T, Mak TW, Sasaki T, Maehama T, MoriM, Suzuki A
  • Journal Title: Nat Med Volume: 17 Pages: 944-951
  • Peer Reviewed
• [Journal Article] In vitro generation of HSC-like cells from murine ESCs/ iPSCs by enforced expression of LIM-homeobox transcription factor Lhx2 (2011)
  • Author(s): Kitajima K, Minehata K, Sakimura K, Nakano T, Hara T
  • Journal Title: Blood Volume: 117 Pages: 3748-3758
  • Peer Reviewed
• [Journal Article] Role for piRNAs and a novel 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 PJ, Toyoda A, Gotoh K, Hiura H, Arima T, Fujiyama A, Sado T, Shibata T, Nakano T, Lin H, Ichiyanagi K, Soloway PD, Sasaki H
  • Journal Title: Science Volume: 332 Pages: 848-852
  • Peer Reviewed
• [Journal Article] Induction of pluripotency in primordial germ cells (2011)
  • Author(s): Kimura T, Nakano T
  • Journal Title: Histol Histopathol Volume: 26 Pages: 643-650
  • Peer Reviewed
• [Journal Article] Sox9 sustains chondrocyte survival and hypertrophy in part through Pik3ca-Akt pathways (2011)
  • Author(s): Ikegami D, Akiyama H, Suzuki A, Nakamura T, Nakano T, Yoshikawa H, Tsumaki N
  • Journal Title: Development Volume: 138 Pages: 1507-1519
  • 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, Chuma S, Yamamoto Y, Kuramochi-Miyagawa S, Totoki Y, Toyoda A, Hoki Y, Fujiyama A, Shibata T, Sado T, Noce T, Nakano T, Nakatsuji N, Lin H, Sasaki H
  • Journal Title: Dev Cell Volume: 20 Pages: 364-375
  • Peer Reviewed
• [Journal Article] 5-hydroxymethylcytosine in the mammalian zygote is linked with epigenetic reprogramming (2011)
  • Author(s): Wossidlo M, Nakamura T, Lepikhov K, Marques J, Zakhartchenko V, Boiani M, Arand J, Nakano T, Reik W, Walter J
  • Journal Title: Nature Commun Volume: 2 Pages: 241-241
  • Peer Reviewed
• [Journal Article] Loss of Lhx1 activity impacts on the localization of primordial germ cells in the mouse (2011)
  • Author(s): Tanaka SS, Yamaguchi YL, Steiner KA, Nakano T, Nishinakamura R, Kwan KM, Behringer RR, Tam PP
  • Journal Title: Dev Dyn Volume: 239 Pages: 2851-9
  • Peer Reviewed
• [Journal Article] 5-hydroxymethylcy tosine in the mammalian zygote is linked with epigenetic reprogramming (2011)
  • Author(s): Wossidlo M, et al
  • Journal Title: Nature Commun Volume: 2 Issue: 1 Pages: 241-241
  • DOI: 10.1038/ncomms1240
  • Peer Reviewed
• [Journal Article] Tseng CP 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,
  • 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, 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 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, Kimura T, Watanabe-Kushima S, Shinohara T, Nakano T
  • Journal Title: Biochem Biophys Res Commun Volume: 392 Pages: 311-316
  • 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] 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 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, Toyoda S, Kuramochi-Miyagawa S, Miyazaki T, Miyazaki S, Tashiro F, Yamato E, Nakano T, Miyazaki J
  • Journal Title: Dev Biol Volume: 335 Pages: 216-217
  • 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, Kuramochi-Miyagawa S, Chuma S, Tanaka T, Nakatsuji N, Kimura T, Nakano T
  • Journal Title: Genes Cells Volume: 14 Pages: 1155-1165
  • 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 Volume: 284 Pages: 6507-6519
  • 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: Exp Hematol Volume: 37 Pages: 334-345
  • 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 hematopoictic 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: Experimental Hematology 37 Pages: 334-345
  • Peer Reviewed
• [Journal Article] The TDRD9-MIWI2 complex is essential for piRNA-mediated retrotransposon silenine in the mouse male eermline (2009)
  • Author(s): Shoji M, (13名), Nakano T, (4名)
  • Journal Title: Developmental Cell 17 Pages: 775-787
  • Peer Reviewed
• [Journal Article] Gtsfl/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, (6名), Nakano T, Miyazaki J
  • Journal Title: Developmental Biology 335 Pages: 216-227
  • Peer Reviewed
• [Journal Article] MILL, a piRNA binding protein, is required for germline stem cell self-renewal and annears 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] 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] DNA Methylation and Hydroxymethylation (2011)
  • Author(s): Nakano T
  • Organizer: MBJS 2011
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2011-12-16
• [Presentation] DNA Methylation and Hydroxymethylation (2011)
  • Author(s): 仲野徹
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 横浜(招待講演)
  • Year and Date: 2011-12-16
• [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(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(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(招待講演)
  • Year and Date: 2010-09-06
• [Presentation] Epigenetic Gene Regulation and GATA factors (2009)
  • Author(s): Nakano T
  • Organizer: Tohoku University Global COE 1st International Sympsium
  • Place of Presentation: Sendai, Japan
  • Year and Date: 2009-12-08
• [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): Nakano T
  • Organizer: International Society of Experimental Hematology
  • Place of Presentation: Athens, Greek
  • 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): Nakano T
  • Organizer: Epigenetics : New Horizons in Japan and Scandinavia
  • Place of Presentation: Stockholm, Sweeden
  • Year and Date: 2009-09-06
• [Presentation] DNA Methylation in Early Embryogenesis (2009)
  • Author(s): Nakano T
  • Organizer: From Imprinting to the Epigenome
  • Place of Presentation: Cambridge, UK
  • 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