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Visualization of subcellular thermoregulation in live cells using fluorescent coiled-coil proteins

Research Project

Project/Area Number 24655151
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field Chemistry related to living body
Research InstitutionKyoto University

Principal Investigator

KIOYNAKA Shigeki  京都大学, 地球環境学堂, 准教授 (90422980)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywordsサーモセンサー / 蛍光タンパク質 / coiled-coil / tsGFP / TWIX
Research Abstract

In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration localized in different subcellular organelles have been proposed. However, previous studies have failed to observe thermogenesis directly in intact organelles. Here, we have visualized intracellular thermogenesis by using a fluorescent thermosensor, tsGFP. Specific targeting of tsGFPs enables visualization of thermogenesis in discrete organelles, notably mitochondria in brown adipocytes and the endoplasmic reticulum in myotubes. Strikingly, in HeLa cells, targeted tsGFP reveals heterogeneity in mitochondrial thermogenesis that is well correlated with the electrochemical gradient. Thus, tsGFPs directly demonstrate subcellular thermotransduction, and will serve as powerful tools to assess thermogenesis in vivo.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (15 results)

All 2014 2013 2012 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (7 results) Remarks (1 results)

  • [Journal Article] Transnitrosylation directs TRPA1 selectivity in N-nitrosamine activators.2014

    • Author(s)
      Kozai D, Kabasawa Y, Ebert M, Kiyonaka S ; Finnan, Otani Y, Numata T, Takahashi N, Mori Y, Ohwada T.
    • Journal Title

      Molecular Pharmacology

      Volume: 85 Issue: 1 Pages: 175-185

    • DOI

      10.1124/mol.113.088864

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Genetically encoded fluorescent thermosensors visualize subcellular thermoregulation in living cells2013

    • Author(s)
      Shigeki Kiyonaka, Taketoshi Kajimoto, Reiko Sakaguchi, Daisuke Shinmi, Mariko Omatsu-Kanbe, Hiroshi Matsuura, Hiromi Imamura, Takenao Yoshizaki, Itaru Hamachi, Takashi Morii, Yasuo Mori
    • Journal Title

      Nature Methods

      Volume: 10 Issue: 12 Pages: 1232-1238

    • DOI

      10.1038/nmeth.2690

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Nuclear life of the voltage-gated Cacnb4 subunit and its role in gene transcription regulation.2013

    • Author(s)
      Ronjat M, Kiyonaka S, Barbado M, De Waard M, Mori Y
    • Journal Title

      Channels

      Volume: 7 Issue: 2 Pages: 119-125

    • DOI

      10.4161/chan.23895

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Molecular determinants for cardiovascular TRPC6 channel regulation by Ca2+/calmodulin-dependent kinase II2013

    • Author(s)
      Juan Shi, Naomi Geshi, Shinichi Takahashi, Shigeki Kiyonaka, Jun Ichikawa, Yaopeng Hu, Yasuo Mori, Yushi Ito, Ryuji Inoue
    • Journal Title

      Journal of Physiology (London)

      Volume: 591(11) Issue: 11 Pages: 2851-2866

    • DOI

      10.1113/jphysiol.2013.251249

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Cacnb4 directly couples electrical activity to gene expression, a process defective in juvenile epilepsy.2012

    • Author(s)
      Tadmouri A
    • Journal Title

      The EMBO journal

      Volume: 31 Issue: 18 Pages: 3730-3744

    • DOI

      10.1038/emboj.2012.226

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Physical and Functional Interaction of the Active Zone Protein CAST/ERC2 and the β-subunit of the Voltage-dependent Ca2+Channel.2012

    • Author(s)
      Kiyonaka S
    • Journal Title

      J. Biochem

      Volume: 152 Issue: 2 Pages: 149-159

    • DOI

      10.1093/jb/mvs054

    • NAID

      10031166124

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Physical and functional interaction of the active zone protein CAST/ERC2 and the β-subunit of the voltage-dependent Ca(2+) channel.2012

    • Author(s)
      Kiyonaka S
    • Journal Title

      Journal of Biochemistry

      Volume: 152 Pages: 149-159

    • NAID

      10031166124

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] Genetically encoded fluorescent thermosensors visualize subcellular thermoregulation in living cells2014

    • Author(s)
      Shigeki Kiyonaka, Taketoshi Kajimoto, Reiko Sakaguchi, Daisuke Shinmi, Mariko Omatsu-Kanbe, Hiroshi Matsuura, Hiromi Imamura, Takenao Yoshizaki, Itaru Hamachi, Takashi Morii and Yasuo Mori
    • Organizer
      the 58th Annual Meeting of the Biophysical Society
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛍光性サーモセンサーの開発と生体の恒常性を担う熱産生機構の可視化2013

    • Author(s)
      清中茂樹、梶本武利、坂口怜子、森井孝、森泰生
    • Organizer
      生理学研究所研究会「新規シグナル伝達分子とその生理学的可能性」
    • Year and Date
      2013-09-26
    • Related Report
      2013 Final Research Report
  • [Presentation] 神経細胞における内在性グルタミン酸受容体のケミカルラベル2013

    • Author(s)
      若山翔、清中茂樹、浜地格
    • Organizer
      第7回バイオ関連化学シンポジウム
    • Related Report
      2013 Final Research Report
  • [Presentation] 神経細胞における内在性グルタミン酸受容体の蛍光ラベル化法の開発2013

    • Author(s)
      若山翔、清中茂樹、浜地格
    • Organizer
      第86回日本生化学会大会
    • Related Report
      2013 Final Research Report
  • [Presentation] 神経伝達物質受容体のケミカルラベル2013

    • Author(s)
      若山翔、清中茂樹、浜地格
    • Organizer
      日本化学会第93春季年会
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛍光性サーモセンサーの開発と生体の恒常性を担う熱産生機構の可視化2013

    • Author(s)
      清中 茂樹、梶本 武利、坂口 怜子、森井 孝、森 泰生
    • Organizer
      生理学研究所研究会「新規シグナル伝達分子とその生理学的可能性」
    • Place of Presentation
      生理学研究所
    • Related Report
      2013 Annual Research Report
  • [Presentation] 神経伝達物質受容体のケミカルラベル2013

    • Author(s)
      若山 翔
    • Organizer
      日本化学会第93春季年会
    • Place of Presentation
      立命館大学(滋賀県)
    • Related Report
      2012 Research-status Report
  • [Remarks]

    • URL

      http://www.sbchem.kyoto-u.ac.jp/hamachi-lab/

    • Related Report
      2013 Final Research Report

URL: 

Published: 2013-05-31   Modified: 2019-07-29  

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