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Cellular memory depending on phosphorylation states of a signaling protein

Research Project

Project/Area Number 15H02394
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Sako Yasushi  国立研究開発法人理化学研究所, 開拓研究本部, 主任研究員 (20215700)

Co-Investigator(Kenkyū-buntansha) 日比野 佳代  国立遺伝学研究所, 構造遺伝学研究センター, 助教 (40435673)
Research Collaborator Okamoto Kenji  
Project Period (FY) 2015-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥40,170,000 (Direct Cost: ¥30,900,000、Indirect Cost: ¥9,270,000)
Fiscal Year 2018: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2017: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2016: ¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
Fiscal Year 2015: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Keywords細胞情報・動態
Outline of Final Research Achievements

A cytoplasmic serine-threonine kinase RAF is a key regulator of cell fate decision in animal cells. RAF has a open/close structural dynamics which can be detected by using intramolecular FRET of a probe molecule of RAF conjugated with two fluorophores at the each end. We found that average conformation of RAF has a wide single-cell variation and the variation correlates with RAF response after cell stimulation. RAF has a multiple phosphorylation sites, of which combination regulates RAF conformation. Single-molecule FRET imaging of RAF revealed the pathway of conformational change upon cell stimulation. These results suggest that activation of various intracellular kinase molecules of RAF changes the RAF conformation which regulate cellular response to the next round of stimulation.

Academic Significance and Societal Importance of the Research Achievements

RAFは細胞増殖・分化などの運命決定を制御する分子であり、その異常は遺伝病の発症や発癌に関係している。RAFの特徴は活性と関連する極めて多数のリン酸化部位と、開閉の構造ダイナミクスを持つことである。我々は多数のRAF分子の平均構造が細胞毎に異なっており、構造がリン酸化の組み合わせと相関することを発見し、詳細な解析を行った。本研究は、細胞内情報処理蛋白質の多重リン酸化を利用した新たな細胞記憶の形成・保持機構を研究し、その分子機構に由来する細胞個性の発現や遺伝的変異による発病の理解に繋がるものである。

Report

(5 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (48 results)

All 2019 2018 2017 2016 2015 Other

All Int'l Joint Research (4 results) Journal Article (18 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 18 results,  Open Access: 15 results,  Acknowledgement Compliant: 3 results) Presentation (22 results) (of which Int'l Joint Research: 7 results,  Invited: 22 results) Remarks (2 results) Patent(Industrial Property Rights) (2 results) (of which Overseas: 1 results)

  • [Int'l Joint Research] Ulsan University(韓国)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] University Colledge Dublin(アイルランド)

    • Related Report
      2017 Annual Research Report
  • [Int'l Joint Research] McGill University(Canada)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] National University of Singapore(Singapore)

    • Related Report
      2016 Annual Research Report
  • [Journal Article] In-cell single-molecule FRET measurements reveal three conformational state changes in RAF protein2019

    • Author(s)
      Okamoto Kenji、Hibino Kayo、Sako Yasushi
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - General Subjects

      Volume: in press Issue: 2 Pages: 1-42

    • DOI

      10.1016/j.bbagen.2019.04.022

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Inferring a nonlinear biochemical network model from a heterogeneous single-cell time course data2018

    • Author(s)
      Shindo Yuki、Kondo Yohei、Sako Yasushi
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 6790-6790

    • DOI

      10.1038/s41598-018-25064-w

    • NAID

      120006502573

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Automated single-molecule imaging in living cells2018

    • Author(s)
      Yasui Masato、Hiroshima Michio、Kozuka Jun、Sako Yasushi、Ueda Masahiro
    • Journal Title

      Nature Communications

      Volume: 9 Issue: 1 Pages: 1-11

    • DOI

      10.1038/s41467-018-05524-7

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule diffusion-based estimation of ligand effects on G protein?coupled receptors2018

    • Author(s)
      Yanagawa Masataka、Hiroshima Michio、Togashi Yuichi、Abe Mitsuhiro、Yamashita Takahiro、Shichida Yoshinori、Murata Masayuki、Ueda Masahiro、Sako Yasushi
    • Journal Title

      Science Signaling

      Volume: 11 Issue: 548 Pages: 1-16

    • DOI

      10.1126/scisignal.aao1917

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-Molecule F?rster Resonance Energy Transfer Measurement Reveals the Dynamic Partially Ordered Structure of the Epidermal Growth Factor Receptor C-Tail Domain2018

    • Author(s)
      Okamoto Kenji、Sako Yasushi
    • Journal Title

      The Journal of Physical Chemistry B

      Volume: 123 Issue: 3 Pages: 571-581

    • DOI

      10.1021/acs.jpcb.8b10066

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule imaging and manipulation of biomolecular machines and systems2018

    • Author(s)
      Iino Ryota、Iida Tatsuya、Nakamura Akihiko、Saita Ei-ichiro、You Huijuan、Sako Yasushi
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - General Subjects

      Volume: 1862 Issue: 2 Pages: 241-252

    • DOI

      10.1016/j.bbagen.2017.08.008

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Transient Acceleration of Epidermal Growth Factor Receptor Dynamics Produces Higher-Order Signaling Clusters2018

    • Author(s)
      Hiroshima Michio、Pack Chan-gi、Kaizu Kazunari、Takahashi Koichi、Ueda Masahiro、Sako Yasushi
    • Journal Title

      Journal of Molecular Biology

      Volume: 430 Issue: 9 Pages: 1386-1401

    • DOI

      10.1016/j.jmb.2018.02.018

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Lipid-Protein Interplay in Dimerization of Juxtamembrane Domains of Epidermal Growth Factor Receptor2018

    • Author(s)
      Maeda Ryo、Sato Takeshi、Okamoto Kenji、Yanagawa Masataka、Sako Yasushi
    • Journal Title

      Biophysical Journal

      Volume: 114 Issue: 4 Pages: 893-903

    • DOI

      10.1016/j.bpj.2017.12.029

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule fluorescence-based analysis of protein conformation, interaction, and oligomerization in cellular systems2017

    • Author(s)
      Okamoto Kenji、Hiroshima Michio、Sako Yasushi
    • Journal Title

      Biophysical Reviews

      Volume: 10 Issue: 2 Pages: 317-326

    • DOI

      10.1007/s12551-017-0366-3

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Recent advances in FRET for the study of protein interactions and dynamics.2017

    • Author(s)
      Okamoto, K. and Sako, Y.
    • Journal Title

      Curr. Opin. Strct. Biol.

      Volume: 46 Pages: 16-23

    • DOI

      10.1016/j.sbi.2017.03.010

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] A role for anti-viral host defense mechanism in the phylogenetic divergence in baculovirus evolution2016

    • Author(s)
      Nagamine, T. and Sako, Y.
    • Journal Title

      PLoS ONE

      Volume: 11

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A novel spingomyeling/cholesterol specific probe reveals the dynamics of the membrane domain during virus and in Nieman-Pick type C2016

    • Author(s)
      Makino, A., Sako, Y. et al.
    • Journal Title

      FASEB J.

      Volume: 31 Issue: 4 Pages: 1301-1322

    • DOI

      10.1096/fj.201500075r

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cortical polarity of the RING protein PAR-2 is maintained by exchange rate kinetics at the cortical-cytoplasm boundary2016

    • Author(s)
      Arata, Y., Hiroshima, M., Pack, C.-G., Ramanujam, R., Motegi, F., Nakazato, K., Wiseman, P. W., Sawa, H., Kobayashi, T. J., Brandao, H. B., Shibata, T., and Sako, Y.
    • Journal Title

      Cell Rep.

      Volume: 16 Issue: 8 Pages: 2156-2168

    • DOI

      10.1016/j.celrep.2016.07.047

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] 細胞内シグナルのアナログ・デジタル変換2016

    • Author(s)
      新土優樹、小迫英尊、佐甲靖志、高橋恒一
    • Journal Title

      生物物理

      Volume: 56 Pages: 334-336

    • NAID

      130005170660

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Interaction of a Novel Fluorescent GTP Analogue with the small G-protein K-Ras2016

    • Author(s)
      Seigo IWATA, Kaori MASUHARA, Nobuhisa UMEKI, Yasushi SAKO and Shinsaku MARUTA
    • Journal Title

      The Journal of Biochemistry

      Volume: 159 Issue: 1 Pages: 41-48

    • DOI

      10.1093/jb/mvv071

    • NAID

      40020725621

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] State transition analysis of spontaneous branch migration of the Holliday junction by photon-based single-molecule fluorescence resonance energy transfer.2016

    • Author(s)
      Okamoto, K. and Sako, Y.
    • Journal Title

      Biophys. Chem.

      Volume: 209 Pages: 21-27

    • DOI

      10.1016/j.bpc.2015.11.004

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Conversion of graded phosphorylation into switch-like nuclear translocation via autoregulatory mechanisms in ERK signalling.2016

    • Author(s)
      Y. Shindo, K. Iwamoto, K. Mouri, K. Hibino, M. Tomita, H. Kosako, Y. Sako and K. Takahashi.
    • Journal Title

      Nature Communications

      Volume: 7 Issue: 1 Pages: 10485-10485

    • DOI

      10.1038/ncomms10485

    • NAID

      120006535049

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Switching of the positive feedback for RAS activation by a concerted function of SOS membrane association domains.2016

    • Author(s)
      Nakamura, Y., Hibino, K., Yanagida, T., and Sako, Y.
    • Journal Title

      Biophys. Physicobiol.

      Volume: 13 Pages: 1-11

    • NAID

      130005121334

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Presentation] Single-molecule pharmacology of G protein-coupled receptors2019

    • Author(s)
      Sako, Y.
    • Organizer
      1st international symposium on Interdisciplinary Approaches to Integrative Understanding of Biological Signaling Networks
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] In cell automated single-molecule analysis and its extensive applications2018

    • Author(s)
      Hiroshima, M., Yasui, M., Kozuka, J., Sako, Y., and Ueda, M.
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Single-molecule analysis of a cell signaling protein, SOS2018

    • Author(s)
      Nakamura, Y., Hibino, K., Yanagida, T., and Sako, Y.
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Sinlgle-molecular pharmacology of GPCRs2018

    • Author(s)
      Sako, Y.
    • Organizer
      The 3rd Biosignal Research Center International Symposium
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Lateral mobility of GPCRs reflects their activation kinetics2018

    • Author(s)
      Sako, Y.
    • Organizer
      The 91st Annual Meeting of the Japanese Biochemical Society Symposium
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞膜受容体の1分子動態、生理学から薬理学へ2018

    • Author(s)
      佐甲靖志
    • Organizer
      日本顕微鏡学会 第42回関東支部講演会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Lipid-protein cooperativity in the regulation of juxtamembrane domain dimer formation in epidermal growth factor receptor2017

    • Author(s)
      Maeda, R., Sato, T., Okamoto, K., and Sako, Y.
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Single-molecule FRET measurement for EGFR-RAS-MAPK signal transduction pathway2017

    • Author(s)
      Okamoto, K., Maeda, R., and Sako, Y.
    • Organizer
      “Deciphering complex energy landscape and kinetic network from single molecules to cells: a new challenge to make theories meet experiments”
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞内情報伝達のイメージング2017

    • Author(s)
      佐甲靖志
    • Organizer
      理研公開シンポジウム「観る・測る・解く」4次元細胞計測の現状と未来
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 細胞膜受容体の運動・会合体形成による情報伝達2017

    • Author(s)
      佐甲靖志
    • Organizer
      第69回日本細胞生物学会大会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 細胞内の1分子イメージング:生理学から薬理学へ2016

    • Author(s)
      佐甲靖志
    • Organizer
      東京理科大学研究推進機構総合研究院 イメージングフロンティアセンターシンポジウム
    • Place of Presentation
      東京理科大学(野田)
    • Year and Date
      2016-12-10
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Motional dynamics of cell membrane receptors2016

    • Author(s)
      Sako, Y.
    • Organizer
      Pioneering New Fields: Forefront of RIKEN’s Science and Beyond
    • Place of Presentation
      RIKEN (Wako)
    • Year and Date
      2016-11-21
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞膜受容体の1分子可視化2016

    • Author(s)
      佐甲靖志
    • Organizer
      第25回日本バイオイメージング学会学術集会
    • Place of Presentation
      名古屋市立大学(名古屋)
    • Year and Date
      2016-09-05
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 細胞内の確率過程2016

    • Author(s)
      佐甲靖志
    • Organizer
      研究会「異常拡散をめぐる最近の進展」
    • Place of Presentation
      慶應義塾大学(横浜)
    • Year and Date
      2016-09-01
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 増殖因子刺激に対するswitch-likeなERKの核移行様式2016

    • Author(s)
      新土優樹 他
    • Organizer
      第68回日本細胞生物学会大会、日本ケミカルバイオロジー学会合第11回年会合同大会
    • Place of Presentation
      京都テルサ(京都)
    • Year and Date
      2016-06-15
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 1分子計測で見るEGF受容体の細胞膜動態2016

    • Author(s)
      佐甲靖志
    • Organizer
      蛋白研セミナー
    • Place of Presentation
      大阪大学、吹田
    • Year and Date
      2016-03-04
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 細胞内情報処理反応の1分子可視化計測2016

    • Author(s)
      佐甲靖志
    • Organizer
      CBI学会 第370回研究講演会
    • Place of Presentation
      東京大学、東京
    • Year and Date
      2016-03-03
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 細胞膜受容体の動的会合体形成と機能2015

    • Author(s)
      佐甲靖志
    • Organizer
      第13回JCGGシンポジウム
    • Place of Presentation
      ウインク愛知、名古屋
    • Year and Date
      2015-10-01
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Single-molecule analysis of cell signaling reactions2015

    • Author(s)
      佐甲靖志
    • Organizer
      第3回 少数性成物学トレーニングコース
    • Place of Presentation
      大阪大学、吹田
    • Year and Date
      2015-08-01
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 情報伝達蛋白質RAFの疾病変異体のコンフォメーションと機能のイメージング計測2015

    • Author(s)
      日比野佳代、上田昌宏、佐甲靖志
    • Organizer
      第67回 日本細胞生物学会大会
    • Place of Presentation
      タワーホール船堀、東京
    • Year and Date
      2015-07-01
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] RAS-MAPK systemの可視化反応解析 1分子病理・薬理学の可能性2015

    • Author(s)
      佐甲靖志
    • Organizer
      岡山大学大学院薬学研究科セミナー
    • Place of Presentation
      岡山大学、岡山
    • Year and Date
      2015-06-26
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 1分子計測で細胞内反応システムを理解する2015

    • Author(s)
      佐甲靖志
    • Organizer
      第12回 原子・分子・光科学 (AMO) 討論会
    • Place of Presentation
      東京大学、東京
    • Year and Date
      2015-06-20
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Remarks] http://www.riken.go.jp/cell-info/

    • Related Report
      2016 Annual Research Report
  • [Remarks] 佐甲細胞情報研究室Webpage

    • URL

      http://www.riken.go.jp/cell-info/

    • Related Report
      2015 Annual Research Report
  • [Patent(Industrial Property Rights)] Method for evaluating activity of G protein-coupled receptor (GPCR)2018

    • Inventor(s)
      Masataka Yanagawa, et al.
    • Industrial Property Rights Holder
      理化学研究所
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Related Report
      2018 Annual Research Report
    • Overseas
  • [Patent(Industrial Property Rights)] Gタンパク質共役型受容体(GPCR)の活性評価方法2017

    • Inventor(s)
      柳川正隆、佐甲靖志、廣島通夫、安井真人、上田昌宏、冨樫祐一
    • Industrial Property Rights Holder
      柳川正隆、佐甲靖志、廣島通夫、安井真人、上田昌宏、冨樫祐一
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2017-084803
    • Filing Date
      2017
    • Related Report
      2017 Annual Research Report

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Published: 2015-04-16   Modified: 2020-03-30  

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