Identification of the CDKA-target factors that regulate cellular changes during stem cell formation

• Project/Area Number: 15K07119
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant molecular biology/Plant physiology
• Research Institution: National Institute for Basic Biology
• Principal Investigator: Ishikawa Masaki 基礎生物学研究所, 生物進化研究部門, 助教 (00586894)
• Research Collaborator: Hasebe Mitsuyasu ; Murata Takashi ; Fujita Tomomichi
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ヒメツリガネゴケ / 幹細胞 / 細胞周期 / CDKA / 細胞運命転換 / 質量分析 / 幹細胞化

Outline of Final Research Achievements

In the moss Physcomitrella patens, when a leaf is excised from a gametophore and cultivated for a few days on culture medium without phytohormone supplementation, leaf cells facing the cut change into cells that are not distinguishable from chloronema apical cells. During this process, a cell cycle regulator, cyclin-dependent kinase A (CDKA), functions in coordination between cell cycle reactivation and acquisition of new cell-type characteristics. To understand roles of CDKA in the stem cell formation, we performed interactome and phosphoproteomic analyses to identify CDKA-target proteins. As a result, we identified a factor involved in the histone modifications, suggesting that CDKA regulates epigenetic modifications to induce cellular changes. In addition, we developed a sensor to detect the CDKA protein kinase activity at tissue levels.

Academic Significance and Societal Importance of the Research Achievements

細胞周期制御因子であるCDKAが、ヒストン修飾変化を制御している可能性が示唆され、CDKAが細胞周期と細胞運命転換の両方を同時に制御する分子機構の一端が見えてきた。このことは、多細胞体制の形成と維持における細胞増殖と細胞分化の相関関係を規定する分子基盤の統合的な理解に繋がることが期待される。またCDKAのキナーゼ活性をモニターするセンサーの開発は、他のプロテインキナーゼや酵素活性をモニターするバイオセンサーへの開発に繋がることが期待される。

Research Products

• [Journal Article] Single-cell transcriptome analysis of Physcomitrella leaf cells during reprogramming using microcapillary manipulation (2019)
  • Author(s): Minoru Kubo, Tomoaki Nishiyama, Yosuke Tamada, Ryosuke Sano, Masaki Ishikawa, Takashi Murata, Akihiro Imai, Daniel Lang, Taku Demura, Ralf Reski, Mitsuyasu Hasebe
  • Journal Title: Nucleic Acid Research Volume: in press Issue: 9 Pages: 4539-4553
  • DOI: 10.1093/nar/gkz181
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 陸上植物がもつ細胞の分化状態を打破するシステム (2018)
  • Author(s): 石川雅樹, 森下美生, 重信秀治, 長谷部光泰
  • Organizer: 日本植物学会第82回大会
  • Invited
• [Presentation] DNA damage induces cellular reprogramming of differentiated cells to stem cells in the moss Physcomitrella patens (2018)
  • Author(s): Nan Gu, Yosuke Tamada, Akihiro Imai, Gergo Palfalvi, Shuji Shigenobu, Masaki Ishikawa, Chunli Chen, Mitsuyasu Hasebe
  • Organizer: 日本遺伝学会第90回大会 国際シンポジウム
  • Int'l Joint Research / Invited
• [Presentation] RNA-seq data analysis of CDKA double knockout in Physcomitrella patens and investigation into affected potassium transport and photosynthesis pathways (2018)
  • Author(s): Brody Frink, Atsushi Takabayashi, Masaki Ishikawa, Yuzuru Suzuki, Sumio Sagano Ayumi, Tanaka, Mitsuyasu Hasebe, Tomomichi Fujita
  • Organizer: 日本植物学会第82回大会
• [Presentation] Requirement Of Kinase Activity Of CDKA On The Novel Functions In The Moss, Physcomitrella patens (2018)
  • Author(s): Eggie Febrianto Ginanjar, Masaki Ishikawa, Masami Sekine, Natsumi Inoue, Mitsuyasu Hasebe, Ooi-Kock Teh, Tomomichi Fujita
  • Organizer: 第60回 日本植物生理学会年会
• [Remarks] 基礎生物学研究所 生物進化研究部門
  • URL: http://www.nibb.ac.jp/sections/evolutionary_biology_and_biodiversity/hasebe/
• [Remarks] 基礎生物学研究所 生物進化研究部門 長谷部研究室
  • URL: http://www.nibb.ac.jp/evodevo/
• [Remarks] 大学共同利用機関法人 自然科学研究機構 基礎生物学研究所
  • URL: http://www.nibb.ac.jp
• [Remarks] 大学共同利用機関法人 自然科学研究機構 基礎生物学研究所 生物進化研究部門
  • URL: http://www.nibb.ac.jp/evodevo/
• [Remarks] 自然科学研究機構 基礎生物学研究所 生物進化研究部門
  • URL: http://www.nibb.ac.jp/evodevo/