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Analysis of Protein Phosphorylation Networks with Designed Kinase and Proteomics

Research Project

Project/Area Number 20K21272
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 38:Agricultural chemistry and related fields
Research InstitutionNagoya University

Principal Investigator

Nakamichi Norihito  名古屋大学, 生命農学研究科, 教授 (90513440)

Project Period (FY) 2020-07-30 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2021: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Keywords植物 / 遺伝子重複 / キナーゼ / CK1 / CKL / リン酸化 / 近接タンパク質ラベリング / プロテオミクス / 阻害剤
Outline of Research at the Start

我々が開発したユニークな植物のタンパク質リン酸化酵素(カゼインキナーゼ1 :CK1) の阻害剤や, 化合物との結合状態に基づいて人工設計するCK1を利用したプロテオミクス解析によって, 遺伝的重複性が障壁になって不明な点の多いシロイヌナズナのCK1ファミリーの関わる生理現象を明らかにする. 本研究の成果は, 遺伝的重複性が問題となって解析が遅れている植物の多くのキナーゼの機能解明へ向けたプロテオミクス研究のモデル系となる.

Outline of Final Research Achievements

We have previously reported that the CKL (Casein Kinase 1-like) gene family is involved in circadian clock regulation in Arabidopsis thaliana, based on analyses using small molecule inhibitors and multiple gene knockdown. However, it is less known that CKL is involved in physiological phenomena other than the clock. The objective of this study was to identify the phosphorylation substrates by creating a modified CKL and performing proteomics of the plant expressing the modified CKL, and ultimately to clarify the physiological processes in which CKL is involved.
Although we ran into unexpected problems in the preparation of the materials, we were able to overcome some of them, and we expect to identify substrates using these materials.

Academic Significance and Societal Importance of the Research Achievements

植物の発生・生理・代謝などあらゆる生命現象の分子機構は, おもに変異体スクリーニングを起点とする遺伝学と原因遺伝子の解析から進められてきた. 順遺伝学の弱点として遺伝学的冗長性のある遺伝子を生命現象の鍵因子として発見しにくい. 現存する植物は染色体倍加の過程を経ており, 重複機能をもつ遺伝子が多く存在していると考えられる.
本研究は, プロテオミクス解析によってCK1の基質タンパク質およびCK1の関わる生理機能を網羅的に同定することを目指した. このようなアプローチは, 重複性の問題だけでなく, タンパク質翻訳後修飾を俯瞰する上で重要な取り組みとなろう.

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (20 results)

All 2023 2022 2021 2020 Other

All Int'l Joint Research (1 results) Journal Article (8 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 8 results,  Open Access: 6 results) Presentation (11 results) (of which Int'l Joint Research: 2 results,  Invited: 3 results)

  • [Int'l Joint Research] University of Southern Californiau(米国)

    • Related Report
      2021 Research-status Report
  • [Journal Article] Chemical biology to dissect molecular mechanisms underlying plant circadian clocks2022

    • Author(s)
      Nakamichi Norihito、Yamaguchi Junichiro、Sato Ayato、Fujimoto Kazuhiro J.、Ota Eisuke
    • Journal Title

      New Phytologist

      Volume: 235 Issue: 4 Pages: 1336-1343

    • DOI

      10.1111/nph.18298

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Plant clock modifications for adapting flowering time to local environments2022

    • Author(s)
      Maeda Akari E、Nakamichi Norihito
    • Journal Title

      Plant Physiology

      Volume: in press Issue: 2 Pages: 952-967

    • DOI

      10.1093/plphys/kiac107

    • Related Report
      2022 Annual Research Report 2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structure?Function Study of a Novel Inhibitor of Cyclin-Dependent Kinase C in Arabidopsis2022

    • Author(s)
      Saito Ami N、Maeda Akari E、Takahara Tomoaki T、Matsuo Hiromi、Nishina Michiya、Ono Azusa、Shiratake Katsuhiro、Notaguchi Michitaka、Yanai Takeshi、Kinoshita Toshinori、Ota Eisuke、Fujimoto Kazuhiro J、Yamaguchi Junichiro、Nakamichi Norihito
    • Journal Title

      Plant and Cell Physiology

      Volume: 63 Issue: 11 Pages: 1720-1728

    • DOI

      10.1093/pcp/pcac127

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Epidermal CCA1 and PMR5 contribute to nonhost resistance in Arabidopsis2022

    • Author(s)
      1.Nami Maeda, Takaya Noguchi, Norihito Nakamichi, Takamasa Suzuki and Atsushi Ishikawa
    • Journal Title

      Biosci. Biotechnol. Biochem.

      Volume: 86 Issue: 12 Pages: 1623-1630

    • DOI

      10.1093/bbb/zbac160

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phosphorylation of RNA Polymerase II by CDKC;2 Maintains the Arabidopsis Circadian Clock Period2022

    • Author(s)
      Uehara Takahiro N、Nonoyama Takashi、Taki Kyomi、Kuwata Keiko、Sato Ayato、Fujimoto Kazuhiro J、Hirota Tsuyoshi、Matsuo Hiromi、Maeda Akari E、Ono Azusa、Takahara Tomoaki T、Tsutsui Hiroki、Suzuki Takamasa、Yanai Takeshi、Kay Steve A、Itami Kenichiro、Kinoshita Toshinori、Yamaguchi Junichiro、Nakamichi Norihito
    • Journal Title

      Plant and Cell Physiology

      Volume: 63 Issue: 4 Pages: 450-462

    • DOI

      10.1093/pcp/pcac011

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] The singularity response reveals entrainment properties of the plant circadian clock2021

    • Author(s)
      Masuda Kosaku、Tokuda Isao T.、Nakamichi Norihito、Fukuda Hirokazu
    • Journal Title

      Nature Communications

      Volume: 12 Issue: 1 Pages: 864-864

    • DOI

      10.1038/s41467-021-21167-7

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] The Transcriptional Network in the Arabidopsis Circadian Clock System2020

    • Author(s)
      Nakamichi Norihito
    • Journal Title

      Genes

      Volume: 11 Issue: 11 Pages: 1284-1284

    • DOI

      10.3390/genes11111284

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] 有用品種から紐解く植物の概日時計メカニズム2020

    • Author(s)
      中道範人
    • Journal Title

      化学と生物

      Volume: 58 Pages: 646-648

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Presentation] The molecular mechanism of temperature compensation in Arabidopsis2023

    • Author(s)
      Akari Maeda, Hiromi Matsuo, Yoshikatsu Matsubayashi, Toshinori Kinoshita, Norihito Nakamichi
    • Organizer
      第64回日本植物生理学会年会
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Synthetic small molecules modulating plant circadian clock2022

    • Author(s)
      Norihito Nakamichi
    • Organizer
      Society for Research on Biological Rhythms 2022
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] PRR5とTOC1はシロイヌナズナ概日時計の温度補償性に関わる2022

    • Author(s)
      前田明里、松尾宏美、松林嘉克、木下俊則、中道範人
    • Organizer
      第29回日本時間生物学会学術大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 植物時計のE3リガーゼZTLの解析2022

    • Author(s)
      谷川陽奈子、松尾宏美、前田明里、中道範人
    • Organizer
      第29回日本時間生物学会学術大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Analysis of plant clock transcription factor PRR72022

    • Author(s)
      山中佐和子、松尾宏美、中道範人
    • Organizer
      第29回日本時間生物学会学術大会
    • Related Report
      2022 Annual Research Report
  • [Presentation] 時計長周期化化合物と標的タンパクの結合モデル2022

    • Author(s)
      前田明里、松尾宏美、藤本和宏、木下俊則、山口潤一郎、中道範人
    • Organizer
      第63回日本植物生理学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 植物概日時計の温度補償性に関わるタンパク質の量的制御メカニズムの解析2022

    • Author(s)
      前田明里、松尾宏美、木下俊則、中道範人
    • Organizer
      第63回日本植物生理学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] Small-molecule modulators of the plant clock2021

    • Author(s)
      Norihito Nakamichi
    • Organizer
      5th Asia Forum on Chronobiology
    • Related Report
      2021 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] シロイヌナズナの概日時計における周期の温度補償性に関わるタンパク質の量的制御2021

    • Author(s)
      前田明里、松尾宏美、木下俊則、中道範人
    • Organizer
      第28回日本時間生物学会学術大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 時計撹乱化合物の作用機序2021

    • Author(s)
      中道範人, 松尾宏美, 大野梓, 前田明里, 佐藤綾人, 伊丹健一郎, 木下俊則, 山口潤一郎
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Research-status Report
  • [Presentation] シロイヌナズナの概日時計の温度補償性が減衰した変異体について2021

    • Author(s)
      前田明里, 松尾宏美, 木下俊則, 中道範人
    • Organizer
      第62回日本植物生理学会年会
    • Related Report
      2020 Research-status Report

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Published: 2020-08-03   Modified: 2024-01-30  

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