RNAを娘細胞に不均等に分配する二種類のメカニズムの解析

• Project Area: Diversity and asymmetry achieved by RNA program
• Project/Area Number: 21112512
• 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: 西田 宏記 大阪大学, 大学院・理学研究科, 教授 (60192689)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥14,560,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥3,360,000); Fiscal Year 2010: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000); Fiscal Year 2009: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: ホヤ / 胚発生 / 局在RNA / 転写因子 / 発生運命 / 内胚葉 / 非対称細胞分裂 / 中胚葉

Research Abstract

単一の受精卵から様々なタイプの細胞を作り出すために、胚は非対称分裂を繰り返す。このとき、二つの娘細胞は異なった発生運命を獲得するが、この非対称性をもたらす機構として、親細胞内で細胞分裂が始まるまでに何らかの因子が偏っており、その因子が二つの娘細胞に不均等に分配されることが重要である。これらの因子はRNAやタンパク質であると考えられ、これらの情報分子の不均等な分配が起こる。本研究では細胞分裂時における非対称性制御プログラムに関して解析を行うことを目的としている。
脊索動物のホヤの胚では、複数種の母性mRNA(postplasmic RNA)が受精後に卵の後方に局在すること、また、中内胚葉細胞が32細胞期に分裂する際に、中胚葉になる娘細胞にのみWntシグナル依存的にNot遺伝子のmRNAを分配することで中胚葉と内胚葉運命を分離していることが明らかになっている。
そこで、Postplasmic RNAとNot mRNAの細胞内局在をもたらすために必要なRNA結合タンパク質を同定することを試みた。そのために、初期胚から精製したRNAを用いライブラリーを作成し、それを用いてPostplasmic RNAとNot mRNAをベイトとしたYeast Three Hybridスクリーニングを行った。その結果、それぞれに対して複数種の候補タンパク質を得ることができたので、現在、それらについてRNA結合能をプルダウンアッセイ等で検証しているところである。

Research Products

• [Journal Article] Simple procedure for sperm cryopreservation in the larvacean tunicate Oikopleura dioica. (2011)
  • Author(s): Ouchi, K., Nishida, H.
  • Journal Title: Zool.Sci. Volume: 28 Pages: 8-11
  • NAID: 210000169624
  • Peer Reviewed
• [Journal Article] Polo-like kinase 1 is required for localization of PEM protein to the centrosome-attracting body and unequal cleavages in ascidian embryos. (2011)
  • Author(s): Negishi, T., Nishida, H
  • Journal Title: Dev.Growth Differ. Volume: 53 Pages: 76-87
  • Peer Reviewed
• [Journal Article] Spatial and temporal expression of two transcriptional isoforms of Lhx3, a LIM class homeobox gene, during embryogenesis of two phylogenetically remote ascidians, Halocynthia roretzi and Ciona intestinalis. (2010)
  • Author(s): Kobayashi, M., Nishida, H.
  • Journal Title: Gene Expression Patterns Volume: 10 Pages: 98-104
  • Peer Reviewed
• [Journal Article] Dual mechanism controls asymmetric spindle position in ascidian germ cell precursors. (2010)
  • Author(s): Prodon, F., Nishida, H.
  • Journal Title: Development Volume: 344 Pages: 284-292
  • Peer Reviewed
• [Journal Article] Macho-1 regulates unequal cell divisions independently of its function as a muscle determinant. (2010)
  • Author(s): Kumano, G., Nishida, H.
  • Journal Title: Dev.Biol. Volume: 344 Pages: 284-292
  • Peer Reviewed
• [Journal Article] Segregation of germ layer fates by nuclear migration-dependent localization of Not mRNA. (2010)
  • Author(s): Takatori, N., Nishida, H.
  • Journal Title: Dev.Cell Volume: 19 Pages: 589-598
  • Peer Reviewed
• [Journal Article] Plasticity of animal genome architecture unmasked by rapid evolution of a pelagic tunicate. (2010)
  • Author(s): Denoeud, F., Nishida, H.
  • Journal Title: Science Volume: 330 Pages: 1381-1385
  • Peer Reviewed
• [Journal Article] Wnt5 is required for notochord cell intercalation in the ascidian Halocynthia roretzi. (2010)
  • Author(s): Niwano, T.
  • Journal Title: Biol.Cell 101 Pages: 645-659
  • Peer Reviewed
• [Journal Article] Spatial and temporal expression of two transcriptional isoforms of Lhχ3, a LIM class homeobox gene, during embryogenesis of two phylogenetically remote ascidians, Halocynthia roretzi and Ciona intestinalis. (2010)
  • Author(s): Kobayashi, M.
  • Journal Title: Gene Expression Patterns (印刷中)
  • Peer Reviewed
• [Journal Article] Actin microfilaments guide the polarized transport of nuclear pore complexes and the cytoplasmic dispersal of Vasa mRNA during GVBD in the ascidian Halocynthia roretzi. (2009)
  • Author(s): Prodon, F.
  • Journal Title: Dev.Biol. 330 Pages: 377-388
  • Peer Reviewed
• [Journal Article] Patterning of an ascidian embryo along the anterior-posterior axis through spatial regulation of competence and induction ability by maternally localized PEM. (2009)
  • Author(s): Kumano, G.
  • Journal Title: Dev.Biol. 331 Pages: 78-88
  • Peer Reviewed
• [Presentation] Expectation from the community II.Cell Bilogy (2010)
  • Author(s): Hiroki Niehida
  • Organizer: 1st Tunicate Information System Meeting
  • Place of Presentation: Hotel Le Royal(ニース、フランス)(招待請演)
  • Year and Date: 2010-11-11
• [Presentation] オタマボヤにおけるCAB様構造体の解析 (2010)
  • Author(s): 幸西翔平、西田宏記
  • Organizer: 日本動物学会
  • Place of Presentation: 東京大学駒場キャンパス
  • Year and Date: 2010-09-23
• [Presentation] Neurula rotation determines left-right asymmetry in ascidian tadpole Larva. (2010)
  • Author(s): Niahide, K., Nishida, H
  • Organizer: 日本発生生物学会
  • Place of Presentation: 県都国際会議場
  • Year and Date: 2010-06-20
• [Presentation] How do embryos count the cell division rounds in each tissue lineage? : An ascidian case. (2010)
  • Author(s): Hiroki Niahida
  • Organizer: 2nd Joint Meeting of the SFBD and JSDB
  • Place of Presentation: パスツール研究所(パリ、フランス)(招待請演)
  • Year and Date: 2010-05-26
• [Presentation] How do embryos count the cell division rounds in each tissue lineage? : An ascidian case (2010)
  • Author(s): Hiroki Nishida
  • Organizer: Cell Cycle and Development Symposium
  • Place of Presentation: 京都ガーデンパレスホテル(京都)
• [Presentation] マボヤ脊索誘導における応答能消失のメカニズム (2009)
  • Author(s): 橋本秀彦
  • Organizer: 日本発生生物学会
  • Place of Presentation: 朱鷺メッセ(新潟)
• [Presentation] Analysis of the expression of Lhx3 genes in two ascidian species, Halocynthia roretzi and Ciona intestinalis (2009)
  • Author(s): Kobayashi, M.
  • Organizer: 日本発生生物学会
  • Place of Presentation: 朱鷺メッセ(新潟)
• [Presentation] Establishment of embryonic axes and regionalization along the axes : Polarity in organisms and cells (2009)
  • Author(s): Hiroki Nishida
  • Organizer: 5th Interantional Tunicate Meeting
  • Place of Presentation: 沖縄産業支援センター(沖縄)
• [Presentation] Patterning of an acidian embryo along the anterior-posterior axis through spatial regulation of competence and induction ability by PEM (2009)
  • Author(s): Gaku Kumano
  • Organizer: 5th Interantional Tunicate Meeting
  • Place of Presentation: 沖縄産業支援センター(沖縄)
• [Presentation] Wnt-dependent asymmetric partitioning of Not mRNA segregates mesoderm and endoderm fates in ascidian embryos (2009)
  • Author(s): Naohito Takatori
  • Organizer: 5th Interantional Tunicate Meeting
  • Place of Presentation: 沖縄産業支援センター(沖縄)
• [Presentation] Screening for the proteins interacting with PEM sheds light on the Plk1 (2009)
  • Author(s): Takefumi Negishi
  • Organizer: 5th Interantional Tunicate Meeting
  • Place of Presentation: 沖縄産業支援センター(沖縄)
• [Presentation] Molecular basis of attenuation of competence to respond FGF signal in ascidian notochord induction (2009)
  • Author(s): Hidehiko Hashimoto
  • Organizer: 5th Interantional Tunicate Meeting
  • Place of Presentation: 沖縄産業支援センター(沖縄)
• [Presentation] Left-right asymmetry of ascidian larvae is determined by rotation of the whole embryos within the vitelline membrane (2009)
  • Author(s): Hiroki Nishida
  • Organizer: 16th国際発生学会
  • Place of Presentation: エジンバラ国際会議センター(英国、エジンバラ)
• [Presentation] Molecular basis of attenuation of competence to respond FGF signal in ascidian notochord induction (2009)
  • Author(s): Hidehiko Hashimoto
  • Organizer: 16th国際発生学会
  • Place of Presentation: エジンバラ国際会議センター(英国、エジンバラ)
• [Presentation] マボヤ胚発生における発生運命と細胞分裂回数の関係 (2009)
  • Author(s): 藤川徹也
  • Organizer: 日本動物学会
  • Place of Presentation: 静岡大学
• [Presentation] マボヤ初期発生におけるTwist-like遺伝子の発現・機能解析 (2009)
  • Author(s): 児玉仁
  • Organizer: 日本動物学会
  • Place of Presentation: 静岡大学
• [Presentation] オタマボヤにおけるCentrosome-Attracting Body (2009)
  • Author(s): 幸西翔平
  • Organizer: 日本動物学会
  • Place of Presentation: 静岡大学
• [Book] 細胞周期フロンティア (2010)
  • Author(s): 西田宏記, (他73名、共著)
  • Publisher: 共立出版社
• [Book] 研究をささえるモデル生物 (2009)
  • Author(s): 西田宏記, 他59名
  • Total Pages: 262
  • Publisher: 化学同人
• [Remarks]
  • URL: http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/nishida/naiyou.html
• [Remarks]
  • URL: http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/nishida/naiyou.html