染色体上の非コードRNAが動的クロマチン構造を制御する仕組み

• Project Area: Dynamic chromatin structure and function
• Project/Area Number: 16H01309
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: 平岡 泰 大阪大学, 生命機能研究科, 教授 (10359078)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000); Fiscal Year 2017: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2016: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
• Keywords: 染色体 / 細胞

Outline of Annual Research Achievements

分裂酵母の減数分裂において相同染色体が対合する仕組みの研究を行い、非コード RNAが相同染色体を認識し対合を促進する分子基盤の一端を明らかにした。非コードRNAとしてsme2遺伝子から転写されるsme2 RNAに着目し、sme2 RNAと相互作用するタンパク質を探索した。GFP融合タンパク質ライブラリーを用いて、sme2 遺伝子領域に局在するタンパク質を蛍光顕微鏡によるビジュアルスクリーニングを行った結果、10種類のsme2 RNA結合タンパク質を同定した。これらのsme2 RNA結合タンパク質は、染色体上でsme2遺伝子領域を含む3箇所の染色体領域に局在した。sme2遺伝子領域以外の２つの染色体領域を特定するために、生化学的なChIP-seq解析（クロマチン免疫沈降産物に含まれるDNAの塩基配列解析）などを行ったが、染色体領域を特定することができなかった。そこで、分裂酵母ゲノムの150箇所の場所それぞれにlacOリピートを挿入した株を活用し、ビジュアルスクリーニングを行った。これら一群の株においては、lacOリピートが互いに約100kb離れた間隔で、ゲノム全体にわたり挿入されており、lacI-GFPで緑色に染まる。それぞれの株でmCherry を融合したsme2 RNA結合タンパク質（赤色）を観察し、共局在を示すlacO挿入部位を特定する。このビジュアルスクリーニングにより、sme2遺伝子領域を含む3箇所の染色体領域を特定することに成功した。この新たに見つかった領域は、sme2遺伝子領域と同様に、sme2 RNA結合タンパク質に依存して、相同染色体対合の頻度が上昇することがわかった（未発表、投稿準備中）。
また、クロマチン構造形成と機能におけるヒストンバリアントの役割（Yamada et al, Nucleic Acids Research 2018）や非コードRNAの役割（Kajitani et al, Proc Natl Acad Sci USA 2017）、核膜タンパク質の役割（Yang et al, J Biochem 2017; Hirano et al, Genes Cells 2018）を明らかにした。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Lem2 is retained at the nuclear envelope through its interaction with Bqt4 in fission yeast (2018)
  • Author(s): Hirano Yasuhiro、Kinugasa Yasuha、Asakawa Haruhiko、Chikashige Yuji、Obuse Chikashi、Haraguchi Tokuko、Hiraoka Yasushi
  • Journal Title: Genes to cells Volume: 23 Issue: 3 Pages: 122-135
  • DOI: 10.1111/gtc.12557
  • Peer Reviewed / Open Access
• [Journal Article] Estimation of GFP-Nucleoporin Amount Based on Fluorescence Microscopy (2018)
  • Author(s): Asakawa Haruhiko、Hiraoka Yasushi、Haraguchi Tokuko
  • Journal Title: Methods in molecular biology Volume: 1721 Pages: 105-115
  • DOI: 10.1007/978-1-4939-7546-4_10
  • ISBN: 9781493975457, 9781493975464
  • Peer Reviewed
• [Journal Article] The histone variant H2A.Z promotes initiation of meiotic recombination in fission yeast. (2018)
  • Author(s): Yamada S, Kugou K, Ding DQ, Fujita Y, Hiraoka Y, Murakami H, Ohta K, Yamada T.
  • Journal Title: Nucleic Acids Res. Volume: 46 Issue: 2 Pages: 609-620
  • DOI: 10.1093/nar/gkx1110
  • Peer Reviewed / Open Access
• [Journal Article] Ser7 of RNAPII-CTD facilitates heterochromatin formation by linking ncRNA to RNAi (2017)
  • Author(s): Kajitani Takuya、Kato Hiroaki、Chikashige Yuji、Tsutsumi Chihiro、Hiraoka Yasushi、Kimura Hiroshi、Ohkawa Yasuyuki、Obuse Chikashi、Hermand Damien、Murakami Yota
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 114 Issue: 52
  • DOI: 10.1073/pnas.1714579115
  • NAID: 120006539374
  • Peer Reviewed / Open Access
• [Journal Article] Function of nuclear membrane proteins in shaping the nuclear envelope integrity during closed mitosis. (2017)
  • Author(s): Yang H-J, Iwamoto M, Hiraoka Y, Haraguchi T
  • Journal Title: Journal of Biochemistry Volume: 161-1 Issue: 6 Pages: 471-477
  • DOI: 10.1093/jb/mvx020
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Compositionally distinct nuclear pore complexes of functionally distinct dimorphic nuclei in ciliate Tetrahymena. (2017)
  • Author(s): Iwamoto M., Osakada H., Mori C., Fukuda Y., Nagao K., Obuse C., Hiraoka Y., and Haraguchi T.
  • Journal Title: J Cell Sci. Volume: 印刷中 Issue: 10 Pages: 1822-1834
  • DOI: 10.1242/jcs.199398
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Fission yeast APC/C activators Slp1 and Fzr1 sequentially trigger two consecutive nuclear divisions during meiosis. (2017)
  • Author(s): Chikashige Y, Yamane M, Okamasa K, Osakada H, Tsutsumi C, Nagahama Y, Fukuta N, Haraguchi T, Hiraoka Y.
  • Journal Title: FEBS letters. Volume: 591 Issue: 7 Pages: 1029-1040
  • DOI: 10.1002/1873-3468.12612
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Visualization of a Specific Genome Locus by the lacO/LacI-GFP System (2017)
  • Author(s): Ding Da-Qiao、Hiraoka Yasushi
  • Journal Title: Cold Spring Harbor protocols Volume: 2017 Issue: 10 Pages: 236-241
  • DOI: 10.1101/pdb.prot091934
  • Peer Reviewed
• [Journal Article] Microscopic Observation of Living Cells Stained with Fluorescent Probes (2017)
  • Author(s): Asakawa Haruhiko、Ding Da-Qiao、Haraguchi Tokuko、Hiraoka Yasushi
  • Journal Title: Cold Spring Harbor protocols Volume: 2017 Issue: 10 Pages: 230-235
  • DOI: 10.1101/pdb.prot079848
  • Peer Reviewed
• [Journal Article] Spatial organization of the Schizosaccharomyces pombe genome within the nucleus. (2017)
  • Author(s): Matsuda A, Asakawa H, Haraguchi T, Hiraoka Y.
  • Journal Title: Yeast. Volume: 34 Issue: 2 Pages: 55-66
  • DOI: 10.1002/yea.3217
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] In vitro reconstitution and biochemical analyses of theSchizosaccharomyces pombe nucleosome. (2017)
  • Author(s): Masako Koyama, Wataru Nagakura, Hiroki Tanaka, Tomoya Kujirai, Yuji Chikashige, Tokuko Haraguchi, Yasushi Hiraoka, Hitoshi Kurumizaka.
  • Journal Title: Biochemical and biophysical research communications. Volume: 482 Issue: 4 Pages: 896-901
  • DOI: 10.1016/j.bbrc.2016.11.130
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Lipid droplet dynamics during Schizosaccharomyces pombe sporulation and their role in spore survival. (2017)
  • Author(s): Yang HJ, Osakada H, Kojidani T, Haraguchi T, Hiraoka Y.
  • Journal Title: Biology open Volume: 6 Pages: 217-222
  • DOI: 10.1242/bio.022384
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A cohesion-based platform for homologous chromosome pairing in meiosis (2016)
  • Author(s): Da-Qiao Ding, Tokuko Haraguchi, Yasushi Hiraoka
  • Journal Title: Current Genetics Volume: 印刷中 Issue: 3 Pages: 499-502
  • DOI: 10.1007/s00294-016-0570-x
  • Peer Reviewed
• [Journal Article] Inner nuclear membrane protein Lem2 augments heterochromatin formation in response to nutritional conditions. (2016)
  • Author(s): Tange Y, Chikashige Y, Takahata S, Kawakami K, Higashi M, Mori C, Kojidani T, Hirano Y, Asakawa H, Murakami Y, Haraguchi T, Hiraoka Y.
  • Journal Title: Genes to Cells Volume: 21 Issue: 8 Pages: 812-832
  • DOI: 10.1111/gtc.12385
  • NAID: 120005906898
  • Peer Reviewed / Open Access
• [Journal Article] Identification of Conserved MEL-28/ELYS Domains with Essential Roles in Nuclear Assembly and Chromosome Segregation. (2016)
  • Author(s): Gomez-Saldivar G, Fernandez A, Hirano Y, Mauro M, Lai A, Ayuso C, Haraguchi T, Hiraoka Y, Piano F, Askjaer P.
  • Journal Title: PLoS genetics. Volume: 12 Issue: 6 Pages: e1006131-e1006131
  • DOI: 10.1371/journal.pgen.1006131
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Depletion of autophagy receptor p62/SQSTM1 enhances the efficiency of gene delivery in mammalian cells. (2016)
  • Author(s): Tsuchiya M, Ogawa H, Koujin T, Kobayashi S, Mori C, Hiraoka Y, Haraguchi T.
  • Journal Title: FEBS letters. Volume: 590 Issue: 16 Pages: 2671-2680
  • DOI: 10.1002/1873-3468.12262
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A genetically encoded probe for live-cell imaging of H4K20 monomethylation (2016)
  • Author(s): Sato Y., Kujirai T., Arai R., Asakawa H., Ohtsuki C., Horikoshi N., Yamagata K., Ueda J., Nagase T., Haraguchi T., Hiraoka Y., Kimura A., Kurumizaka H., Kimura, H.
  • Journal Title: J. Mol. Biol. Volume: 428 Issue: 20 Pages: 3885-3902
  • DOI: 10.1016/j.jmb.2016.08.010
  • Peer Reviewed / Open Access
• [Journal Article] Borna Disease Virus Assembles Porous Cage-like Viral Factories in the Nucleus. (2016)
  • Author(s): Hirai Y, Hirano Y, Matsuda A, Hiraoka Y, Honda T, Tomonaga K.
  • Journal Title: The Journal of biological chemistry. Volume: 291 Issue: 50 Pages: 25789-25798
  • DOI: 10.1074/jbc.m116.746396
  • Peer Reviewed / Open Access
• [Journal Article] Meiotic cohesion-based chromosome structure is essential for homologous chromosome pairing in Schizosaccharomyces pombe (2015)
  • Author(s): Ding DQ, Matsuda A, Okamasa K, Nagahama Y, Haraguchi T, Hiraoka Y
  • Journal Title: Chromosoma Volume: special issue Issue: 2 Pages: 1-10
  • DOI: 10.1007/s00412-015-0551-8
  • Peer Reviewed / Open Access
• [Presentation] A framework of meiotic chromosomes for homologous pairing. (2017)
  • Author(s): 平岡泰
  • Organizer: 広島大学クロマチン動態数理研究拠点"5th International Symposium of the Mathematics on Chromatin Live Synamics"
  • Place of Presentation: 広島芸術文化ホールくらら（広島県東広島市）
  • Year and Date: 2017-03-07
  • Int'l Joint Research / Invited
• [Presentation] ヒストン不足が引き起こす染色体の不分離 (2017)
  • Author(s): 平岡泰
  • Organizer: よこはまＮＭＲ研究会 第56回ワークショップ「ヌクレオームとビッグデータ」
  • Place of Presentation: 理化学研究所（神奈川県横浜市）
  • Year and Date: 2017-03-03
  • Invited
• [Presentation] Chromatin-anchoring nuclear membrane proteins in fission yeast (2017)
  • Author(s): Hiraoka Y
  • Organizer: FMIシンポジウム
  • Place of Presentation: Friedrich Miescher Institute for Biomedical Research（Basel, Switzerland）
  • Year and Date: 2017-01-20
  • Int'l Joint Research / Invited
• [Presentation] Fluorescence microscopy for studying chromosome dynamics in living cells (2017)
  • Author(s): Hiraoka Yasushi
  • Organizer: (Lecture) National Tsing Hua University
  • Int'l Joint Research / Invited
• [Presentation] RNA transcription and termination factors are important in meiotic homologous chromosome pairing in S. pombe (2017)
  • Author(s): Ding Da-Qiao、Hiraoka Yasushi
  • Organizer: EMBO Conference on Meiosis 2017
  • Int'l Joint Research
• [Presentation] A new method for three-dimensional chromatic correction for fluorescence microscopy (2017)
  • Author(s): Matsuda Atsushi、Haraguchi Tokuko、Hiraoka Yasushi
  • Organizer: The 24th Congress of the International Commission for Optics
  • Int'l Joint Research
• [Presentation] A cohesin-based platform of the chromosome for homologous chromosome pairing in fission yeast meiosis (2017)
  • Author(s): Ding Da-Qiao、Haraguchi Tokuko、Hiraoka Yasushi
  • Organizer: SMC protein
  • Int'l Joint Research / Invited
• [Presentation] 染色体上の非コード RNAが減数分裂期の相同染色体対合を促進する (2017)
  • Author(s): 丁大橋、原口徳子、平岡泰
  • Organizer: 第40回日本分子生物学会年会
• [Presentation] 減数分裂期相同染色体対合における非コードRNA の役割 (2017)
  • Author(s): 丁大橋、原口徳子、平岡泰
  • Organizer: 第35回染色体ワークショップ・第16回核ダイナミクス研究会
• [Presentation] ヒストンH2Aの不足が染色体の不分離を引き起こす (2017)
  • Author(s): 山本孝治、長濵有紀、原口徳子、平岡泰
  • Organizer: 酵母遺伝学フォーラム第50回研究報告会
• [Presentation] Chromosome association of noncoding RNA during homologous chromosome pairing in fission yeast meiosis (2017)
  • Author(s): Ding Da-Qiao、Haraguchi Tokuko、Hiraoka Yasushi
  • Organizer: 第55回日本生物物理学会年会
• [Presentation] Lem2 Localization to the Nuclear Envelope Is Biased by Its Interaction with Another Nuclear Membrane Protein Bqt4 in Fission Yeast (2017)
  • Author(s): Hirano Yasuhiro、Kinugasa Yasuha、Obuse Chikashi、Asakawa Haruhiko、Chikashige Yuji、Haraguchi Tokuko、Hiraoka Yasushi
  • Organizer: 9th International Fission Yeast Meeting
  • Int'l Joint Research
• [Presentation] Organization of The Nuclear Pore Core Complex Revealed by Immunoelectron Microscopy in The Fission Yeast Schizosaccharomyces pombe (2017)
  • Author(s): Asakawa Haruhiko、Kojidani Tomoko、Yang Hui-Ju、Osakada Hiroko、Nagao Koji、Obuse Chikashi、Hiraoka Yasushi、Haraguchi Tokuko
  • Organizer: 9th International Fission Yeast Meeting
  • Int'l Joint Research
• [Presentation] A framework of meiotic chromosomes for homologous pairing. (2016)
  • Author(s): Hiraoka Y
  • Organizer: 2016 Workshop on the Molecular and Physical Biology of Chromosomes
  • Place of Presentation: Woods Hole Marine Biological Laboratory（Massachusetts, USA）
  • Year and Date: 2016-09-11
  • Int'l Joint Research / Invited
• [Presentation] 遺伝子のすがたを生きたまま見る (2016)
  • Author(s): 平岡泰
  • Organizer: 一般公開シンポジウム「遺伝子のすがた -カラダの中でおこる不思議-」
  • Place of Presentation: 早稲田大学国際会議場井深大記念ホール（東京都新宿区）
  • Year and Date: 2016-08-21
  • Invited
• [Book] Fission Yeast: A Laboratory Manual (2016)
  • Author(s): Ding DQ, Hiraoka Y
  • Total Pages: 487
  • Publisher: Cold Spring Harbor Laboratory Press
• [Book] Fission Yeast: A Laboratory Manual (2016)
  • Author(s): Asakawa H, Ding DQ, Haraguchi T, Hiraoka Y
  • Total Pages: 487
  • Publisher: Cold Spring Harbor Laboratory Press
• [Remarks] 細胞核ダイナミクス研究室
  • URL: http://www.fbs.osaka-u.ac.jp/jpn/general/lab/07/
• [Remarks] 細胞核ダイナミクス研究室 研究成果
  • URL: http://www.fbs.osaka-u.ac.jp/jpn/general/lab/07/result/
• [Remarks] 新学術領域研究「動的クロマチン構造と機能」
  • URL: http://nucleosome.kyushu-u.ac.jp