Isoform selectivity of circadian clock proteins CRY1 and CRY2 analyzed by new chemical tools

• Project/Area Number: 18H02402
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43030:Functional biochemistry-related
• Research Institution: Nagoya University
• Principal Investigator: Hirota Tsuyoshi 名古屋大学, トランスフォーマティブ生命分子研究所, 特任准教授 (50372412)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2018: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
• Keywords: 概日時計 / ケミカルバイオロジー / 化合物 / CRYタンパク質 / 選択性

Outline of Final Research Achievements

CRY proteins constituting the mammalian circadian clock have two closely related homologs, CRY1 and CRY2. Functional differences of these proteins provide a model system to tackle fundamental questions regarding how the circadian clock precisely oscillates and controls physiological outputs. In this study, we discovered isoform-selective compounds against CRY1 and CRY2, and by analyzing their mechanism of action, revealed molecular mechanism underlying difference of CRY1 and CRY2.

Academic Significance and Societal Importance of the Research Achievements

CRY1とCRY2は配列が非常によく似ているため、選択的な化合物のデザインは困難であった。本研究では、配列がほぼ同一である化合物結合ポケットに対して選択性を生み出すユニークな化合物を見出し、それらの作用機構を解明した。概日時計が乱れるリズム障害は現代社会における深刻な問題となっており、本研究による制御機構の理解と特異的な調節ツールの開発は、CRYを利用した疾患治療法の開発に向けた重要な足がかりとなるであろう。

Research Products

• [Int'l Joint Research] University of Southern California(米国)
• [Int'l Joint Research] University of Groningen(オランダ)
• [Int'l Joint Research] University of Padova(イタリア)
• [Int'l Joint Research] University of Southern California/University of California Santa Cruz/California Institute of Technology(米国)
• [Int'l Joint Research] University of Groningen(オランダ)
• [Int'l Joint Research] University of Southern California/The Scripps Research Institute(米国)
• [Int'l Joint Research] Korea Inst Sci Tech(韓国)
• [Journal Article] Reductive stability evaluation of 6-azopurine photoswitches for the regulation of CKIα activity and circadian rhythms (2021)
  • Author(s): Kolarski Dusan、Sugiyama Akiko、Rodat Theo、Schulte Albert、Peifer Christian、Itami Kenichiro、Hirota Tsuyoshi、Feringa Ben L.、Szymanski Wiktor
  • Journal Title: Organic & Biomolecular Chemistry Volume: 19 Issue: 10 Pages: 2312-2321
  • DOI: 10.1039/d1ob00014d
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparing the efficacy and selectivity of Ck2 inhibitors. A phosphoproteomics approach (2021)
  • Author(s): Borgo Christian、Cesaro Luca、Hirota Tsuyoshi、Kuwata Keiko、D’Amore Claudio、Ruppert Thomas、Blatnik Renata、Salvi Mauro、Pinna Lorenzo A.
  • Journal Title: European Journal of Medicinal Chemistry Volume: 214 Pages: 113217-113217
  • DOI: 10.1016/j.ejmech.2021.113217
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Photopharmacological Manipulation of Mammalian CRY1 for Regulation of the Circadian Clock (2021)
  • Author(s): Kolarski Dusan、Miller Simon、Oshima Tsuyoshi、Nagai Yoshiko、Aoki Yugo、Kobauri Piermichele、Srivastava Ashutosh、Sugiyama Akiko、Amaike Kazuma、Sato Ayato、Tama Florence、Szymanski Wiktor、Feringa Ben L.、Itami Kenichiro、Hirota Tsuyoshi
  • Journal Title: Journal of the American Chemical Society Volume: 143 Issue: 4 Pages: 2078-2087
  • DOI: 10.1021/jacs.0c12280
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A N-terminally deleted form of the CK2α’ catalytic subunit is sufficient to support cell viability (2020)
  • Author(s): Borgo Christian、D’Amore Claudio、Cesaro Luca、Itami Kenichiro、Hirota Tsuyoshi、Salvi Mauro、Pinna Lorenzo A.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 531 Issue: 3 Pages: 409-415
  • DOI: 10.1016/j.bbrc.2020.07.112
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] “Janus” efficacy of CX-5011: CK2 inhibition and methuosis induction by independent mechanisms (2020)
  • Author(s): D'Amore Claudio、Moro Enrico、Borgo Christian、Itami Kenichiro、Hirota Tsuyoshi、Pinna Lorenzo A.、Salvi Mauro
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Molecular Cell Research Volume: 1867 Issue: 11 Pages: 118807-118807
  • DOI: 10.1016/j.bbamcr.2020.118807
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Small Molecules Modulating Mammalian Biological Clocks: Exciting New Opportunities for Synthetic Chemistry (2020)
  • Author(s): Amaike Kazuma、Oshima Tsuyoshi、Skoulding Nicola Stephanie、Toyama Yoshifumi、Hirota Tsuyoshi、Itami Kenichiro
  • Journal Title: Chem Volume: 6 Issue: 9 Pages: 2186-2198
  • DOI: 10.1016/j.chempr.2020.08.011
  • Peer Reviewed
• [Journal Article] An Isoform-Selective Modulator of Cryptochrome 1 Regulates Circadian Rhythms in Mammals (2020)
  • Author(s): Miller Simon、Aikawa Yoshiki、Sugiyama Akiko、Nagai Yoshiko、Hara Aya、Oshima Tsuyoshi、Amaike Kazuma、Kay Steve A.、Itami Kenichiro、Hirota Tsuyoshi
  • Journal Title: Cell Chemical Biology Volume: 27 Issue: 9 Pages: 1192-1198
  • DOI: 10.1016/j.chembiol.2020.05.008
  • NAID: 120006957723
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Isoform-selective regulation of mammalian cryptochromes (2020)
  • Author(s): Miller Simon、Son You Lee、Aikawa Yoshiki、Makino Eri、Nagai Yoshiko、Srivastava Ashutosh、Oshima Tsuyoshi、Sugiyama Akiko、Hara Aya、Abe Kazuhiro、Hirata Kunio、Oishi Shinya、Hagihara Shinya、Sato Ayato、Tama Florence、Itami Kenichiro、Kay Steve A.、Hatori Megumi、Hirota Tsuyoshi
  • Journal Title: Nature Chemical Biology Volume: - Issue: 6 Pages: 676-685
  • DOI: 10.1038/s41589-020-0505-1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Pharmacological Interventions to Circadian Clocks and Their Molecular Bases (2020)
  • Author(s): Miller Simon、Hirota Tsuyoshi
  • Journal Title: Journal of Molecular Biology Volume: - Issue: 12 Pages: 3498-3514
  • DOI: 10.1016/j.jmb.2020.01.003
  • Peer Reviewed / Open Access
• [Journal Article] 化合物を使って体内時計を操る (2020)
  • Author(s): 松田 智宏、廣田 毅
  • Journal Title: 化学 Volume: 75 Pages: 66-67
• [Journal Article] Dynamics at the serine loop underlie differential affinity of cryptochromes for CLOCK:BMAL1 to control circadian timing (2020)
  • Author(s): Fribourgh Jennifer L、Srivastava Ashutosh、Sandate Colby R、Michael Alicia K、Hsu Peter L、Rakers Christin、Nguyen Leslee T、Torgrimson Megan R、Parico Gian Carlo G、Tripathi Sarvind、Zheng Ning、Lander Gabriel C、Hirota Tsuyoshi、Tama Florence、Partch Carrie L
  • Journal Title: eLife Volume: 9
  • DOI: 10.7554/elife.55275
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Controlling the Circadian Clock with High Temporal Resolution through Photodosing (2019)
  • Author(s): Kolarski Dusan、Sugiyama Akiko、Breton Ghislain、Rakers Christin、Ono Daisuke、Schulte Albert、Tama Florence、Itami Kenichiro、Szymanski Wiktor、Hirota Tsuyoshi、Feringa Ben L.
  • Journal Title: Journal of the American Chemical Society Volume: 141 Issue: 40 Pages: 15784-15791
  • DOI: 10.1021/jacs.9b05445
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of pathways that regulate circadian rhythms using a larval zebrafish small molecule screen (2019)
  • Author(s): Mosser Eric A.、Chiu Cindy N.、Tamai T. Katherine、Hirota Tsuyoshi、Li Suna、Hui May、Wang Amy、Singh Chanpreet、Giovanni Andrew、Kay Steve A.、Prober David A.
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 12405-12405
  • DOI: 10.1038/s41598-019-48914-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chemical Synthesis of Atomically Tailored SUMO E2 Conjugating Enzymes for the Formation of Covalently Linked SUMO?E2?E3 Ligase Ternary Complexes (2019)
  • Author(s): Zhang Yinfeng、Hirota Tsuyoshi、Kuwata Keiko、Oishi Shunsuke、Gramani Subramanian G.、Bode Jeffrey W.
  • Journal Title: Journal of the American Chemical Society Volume: 141 Issue: 37 Pages: 14742-14751
  • DOI: 10.1021/jacs.9b06820
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Casein kinase 1 family regulates PRR5 and TOC1 in the Arabidopsis circadian clock (2019)
  • Author(s): Uehara TN.、Mizutani Y、Kuwata K、Hirota T、Sato A、Mizoi J、Takao S、Matsuo H、Suzuki T、Ito S、Saito AN.、Nishiwaki-Ohkawa T、Yamaguchi-Shinozaki K、Yoshimura T、Kay SA.、Itami K、Kinoshita T、Yamaguchi J、Nakamichi N.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Issue: 23 Pages: 11528-11536
  • DOI: 10.1073/pnas.1903357116
  • NAID: 120006645300
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell-based screen identifies a new potent and highly selective CK2 inhibitor for modulation of circadian rhythms and cancer cell growth. (2019)
  • Author(s): Oshima T, Niwa Y, Kuwata K, Srivastava A, Hyoda T, Tsuchiya Y, Kumagai M, Tsuyuguchi M, Tamaru T, Sugiyama A, Ono N, Zolboot N, Aikawa Y, Oishi S, Nonami A, Arai F, Hagihara S, Yamaguchi J, Tama F, Kunisaki Y, Yagita K, Ikeda M, Kinoshita T, Kay SA, Itami K, Hirota T.
  • Journal Title: Science Advance Volume: 23 Issue: 1 Pages: 9060-9060
  • DOI: 10.1126/sciadv.aau9060
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chemical Control of Mammalian Circadian Behavior through Dual Inhibition of Casein Kinase Iα and δ (2019)
  • Author(s): Lee Jae Wook、Hirota Tsuyoshi、Ono Daisuke、Honma Sato、Honma Ken-ichi、Park Keunwan、Kay Steve A.
  • Journal Title: Journal of Medicinal Chemistry Volume: 62 Issue: 4 Pages: 1989-1998
  • DOI: 10.1021/acs.jmedchem.8b01541
  • NAID: 120006645104
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 時計遺伝子から考える脳機能と食 (2019)
  • Author(s): 羽鳥 恵、廣田 毅
  • Journal Title: FOOD Style21 Volume: 23 Pages: 80-83
• [Journal Article] Nuclear receptor HNF4A transrepresses CLOCK:BMAL1 and modulates tissue-specific circadian networks (2018)
  • Author(s): Qu Meng、Duffy Tomas、Hirota Tsuyoshi、Kay Steve A.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 115 Issue: 52
  • DOI: 10.1073/pnas.1816411115
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Reversible Control of the Circadian Period by Photoswitchable Longdaysin (2020)
  • Author(s): Tsuyoshi Hirota, Dusan Kolarski, Akiko Sugiyama, Wiktor Szymanski, and Ben L. Feringa
  • Organizer: Socirty for Research on Biological Rhythms 2020 Meeting
  • Int'l Joint Research
• [Presentation] 時計タンパク質に作用する低分子化合物の開発 (2020)
  • Author(s): 廣田 毅
  • Organizer: 第41回日本臨床薬理学会学術総会
  • Invited
• [Presentation] Small molecule modulators of the circadian clock function (2020)
  • Author(s): Tsuyoshi Hirota
  • Organizer: 日本薬理学会 第93回年会
  • Invited
• [Presentation] 哺乳類CRYタンパク質のアイソフォーム選択的な制御 (2019)
  • Author(s): 廣田 毅
  • Organizer: 第2回構造生命科学研究会
• [Presentation] 概日時計タンパク質を標的とする低分子化合物 (2019)
  • Author(s): 廣田 毅
  • Organizer: 2019年度BINDS公開シンポジウム
  • Invited
• [Presentation] Chemical and structural biology approach for circadian biochemistry of clock proteins (2019)
  • Author(s): Tsuyoshi Hirota
  • Organizer: 第92回日本生化学会大会
  • Invited
• [Presentation] Small molecule modulators of mammalian clock proteins (2019)
  • Author(s): Tsuyoshi Hirota
  • Organizer: 2019 CIBR Beijing Conference on Brain & Behavior
  • Int'l Joint Research / Invited
• [Presentation] 体内時計を調節する化合物の探索 (2019)
  • Author(s): 廣田 毅
  • Organizer: 日本農芸化学会2019年度大会
  • Invited
• [Presentation] 構造生物学とケミカルバイオロジーの融合による概日時計の創薬研究 (2019)
  • Author(s): 廣田 毅
  • Organizer: 蛋白研セミナー 第1回構造生命科学研究会
  • Invited
• [Presentation] 新規の概日リズム調節化合物の同定とがん細胞増殖に対する効果 (2019)
  • Author(s): 廣田 毅
  • Organizer: 第2回がんと代謝研究会若手の会
• [Presentation] Dissecting mammalian clock mechanism with chemical tools (2018)
  • Author(s): Hirota, T.
  • Organizer: International Symposium on Biological Rhythms
  • Int'l Joint Research / Invited
• [Presentation] ケミカルバイオロジーを応用した哺乳類の概日時計のメカニズム解析 (2018)
  • Author(s): 廣田 毅
  • Organizer: 第3回先端ケミカルバイオロジー研究会
  • Invited
• [Presentation] Identification of CRY1/CRY2 selective compounds (2018)
  • Author(s): Hirota, T.
  • Organizer: 2018 Society for Research on Biological Rhythm Meeting
  • Int'l Joint Research
• [Remarks] 名古屋大学トランスフォーマティブ生命分子研究所Kay-廣田グループ
  • URL: http://www.itbm.nagoya-u.ac.jp/ja/kay-hirota_group/