Photoregulation of gene expression and enzymatic activity by dimerization of LOV protein

• Project/Area Number: 26440077
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: Hisatomi Osamu 大阪大学, 理学研究科, 准教授 (60231544)
• Research Collaborator: IWASA Tatsuo ; HIDEKI Kandori ; SAKAI Takaomi ; SAKAKIBARA Shunsuke ; SHICHIDA Yoshinori ; TERAZIMA Masahide ; NAKAMURA Ryousuke ; HAMADA Norio ; FUKADA Yoshitaka ; MARUTA Shinsaku ; MINO Hiroyuki ; YAMASHITA Hayato
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2014: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 光制御 / 転写因子 / 光遺伝学 / LOVドメイン / bZIPドメイン / 光スイッチ / 分子間相互作用

Outline of Final Research Achievements

Photozipper (PZ) is a protein module consisting of a bZIP and a LOV domain of AUREO1. We improved measurement systems of biomolecular complexes and investigated PZ. PZ is monomeric in the dark, and blue light induces dimerization of PZ, which subsequently increases its affinity for the target DNA sequence. PZ is, therefore, assumed to be a light-activated bZIP module. Then, we found that an intramolecular interaction between ZIP and LOV probably stabilize monomeric form in the dark state, and that intermolecular interactions of ZIP-ZIP and LOV-LOV stabilizes dimeric form in the light state. PZ fused with fluorescent proteins underwent dimerization and increased its affinity for the target DNA sequence upon illumination. Our results suggested that PZ provide a new approach for regulating the gene expression and enzymatic activities.

Research Products

• [Journal Article] Time-resolved Detection of Light-induced Dimerization of Monomeric Aureochrome-1 and Change in Affinity for DNA (2016)
  • Author(s): Y. Akiyama, Y. Nakasone, Y. Nakatani, O. Hisatomi, M. Terazima
  • Journal Title: J. Phys. Chem. B Volume: 120 Issue: 30 Pages: 7360-7370
  • DOI: 10.1021/acs.jpcb.6b05760
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Molecular mechanism of Photozipper, a light-regulated dimerizing module consisting of the bZIP and LOV domains of Aureochrome-1 (2015)
  • Author(s): Y. Nakatani, O. Hisatomi
  • Journal Title: Biochemistry Volume: E-pub Issue: 21 Pages: 10-10
  • DOI: 10.1021/acs.biochem.5b00320
  • Peer Reviewed
• [Journal Article] A light-regulated bZIP module, Photozipper, induces the binding of fused proteins to the target DNA sequence in a blue light-dependent manner. (2015)
  • Author(s): Osamu Hisatomi, Keigo Furuya
  • Journal Title: Photochem. Photobiol. Sci. Volume: 14 Issue: 11 Pages: 1998-2006
  • DOI: 10.1039/c5pp00178a
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Accuracy of protein size estimates based on light scattering measurements. (2014)
  • Author(s): Takeuchi, K., Nakatani, Y. and Hisatomi, O.
  • Journal Title: Open J. Biophys. Volume: 4 Issue: 02 Pages: 83-91
  • DOI: 10.4236/ojbiphy.2014.42009
  • Peer Reviewed / Open Access
• [Journal Article] Blue light-induced dimerization of monomeric aureochrome-1 enhances its affinity for the target sequence. (2014)
  • Author(s): O. Hisatomi, Y. Nakatani, K. Takahashi, H. Kataoka
  • Journal Title: J. Biol. Chem. Volume: 289 Issue: 25 Pages: 17379-17391
  • DOI: 10.1074/jbc.m114.554618
  • Peer Reviewed
• [Journal Article] Molecular Mechanism of a Photoregulated bZIP Transcription Factor and Its Application as a Molecular Tool (2014)
  • Author(s): 久冨 修
  • Journal Title: Seibutsu Butsuri Volume: 54 Issue: 6 Pages: 307-310
  • DOI: 10.2142/biophys.54.307
  • NAID: 130004713030
  • ISSN: 0582-4052, 1347-4219
  • Peer Reviewed
• [Presentation] コアフコース認識レクチンPhoSLの構造解析 (2017)
  • Author(s): 井上萌恵、Nguyen Minh Hien、和泉雅之、岡本亮、小林夕香、上野泰、岡本裕樹、林文晶、久冨修、梶原康宏
  • Organizer: 日本化学会 第97春季年会
  • Place of Presentation: 横浜、神奈川
  • Year and Date: 2017-03-16
• [Presentation] Reaction mechanism in Photozipper monitored by Electron Paramagntic Resonanced (2016)
  • Author(s): K. Ozeki, H. Nagashima, O. Hisatomi, H. Mino
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-25
• [Presentation] The quantitative analyses for the DNA binding of Photozipper (2016)
  • Author(s): Y. Nakatani1, O. Hisatomi
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-25
• [Presentation] The DNA-binding of a light-regulated bZIP module, photozipper, analyzed by quartz crystal microbalance (2016)
  • Author(s): S. Tateyama, O. Hisatomi
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-25
• [Presentation] Mutational analyses of the conformational switching of a light-regulated bZIP module, Photozipper (2016)
  • Author(s): O. Hisatomi
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-25
• [Presentation] Thermal activation rates of visual pigments expressed in rods (2016)
  • Author(s): K. Kojima, Y. Matsutani, M. Yanagawa, T. Yamashita, Y. Imamoto, O. Hisatomi, Y. Yamano, A. Wada, and Y. Shichida
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-25
• [Presentation] Optimization mechanism of vertebrate visual pigments for photic environment (2016)
  • Author(s): Y. Matsutani, K. Kojima, M. Yanagawa, T. Yamashita, Y. Imamoto, O. Hisatomi, Y. Yamano, A. Wada, Y. Shichida
  • Organizer: 日本動物学会第 87回大会
  • Place of Presentation: 沖縄
  • Year and Date: 2016-11-14
• [Presentation] 機能性タンパク質photozipperの反応過程の解析 (2016)
  • Author(s): 小関康平、長嶋宏樹、久冨修、三野広幸
  • Organizer: 第55回電子スピンサイエンス学会年会（SEST2016)
  • Place of Presentation: 大阪市大、大阪
  • Year and Date: 2016-11-10
• [Presentation] Thermal isomerization rate of retinal chromophore in blue-sensitive cone pigments expressed in amphibian green rods (2016)
  • Author(s): K. Kojima, Y. Matsutani, T. Yamashita, M. Yanagawa, Y. Imamoto, Y. Yamano, A. Wada, O. Hisatomi, Y. Shichida
  • Organizer: 16th International Conference on Retinal ProteinsI (ICRP2016)
  • Place of Presentation: Potsdam, Germany
  • Year and Date: 2016-10-02
  • Int'l Joint Research / Invited
• [Presentation] DNA-binding of a light-regulated bZIP module, Photozipper (2016)
  • Author(s): Y. Nakatani and O. Hisatomi
  • Organizer: 第38回日本比較生理生化学会、東京大会（jscpb2016）
  • Place of Presentation: 玉川大学、東京
  • Year and Date: 2016-09-02
• [Presentation] 光制御型 bZIP モジュールの機能解析 (2016)
  • Author(s): 久冨修、中谷陽一、竹内健
  • Organizer: 第19回 日本光生物学協会 年会
  • Place of Presentation: 東京大学、東京
  • Year and Date: 2016-07-28
• [Presentation] 錐体視物質の暗環境適応 (2016)
  • Author(s): 松谷優樹、小島慧一、柳川正隆、山下高廣、今元泰、久富修、山野由美子、和田昭盛、七田芳則
  • Organizer: 第19回 日本光生物学協会 年会
  • Place of Presentation: 東京大学、東京
  • Year and Date: 2016-07-28
• [Presentation] Undergraduate education in geochemistry and cosmochemistry at Osaka University, Japan (2016)
  • Author(s): H. Yabuta, T. Kondo, S. Nakashima, S. Sasaki, H. Shibai, K. Terada, M. Toyoda, T. HironoI, O. Hisatomi, K. Saiki, H. Terasaki, C. Uyeda, C. Yamanaka, J. Aoki, K. Hashizume, Y. Kawai, T. Sakaiya, A. Tani
  • Organizer: Goldschmidt Conference
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2016-06-26
  • Int'l Joint Research
• [Presentation] Blue light-induced DNA binding of an engineered bZIP module, Photozipper (2016)
  • Author(s): O. Hisatomi
  • Organizer: 4th Awaji International Workshop on “Electron Spin Science & Technology: Biological and Materials Science Oriented Applications” (AWEST 2016)
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2016-06-19
  • Int'l Joint Research / Invited
• [Presentation] Molecular mechanisms of a light-activatable bZIP module, Photozipper (2016)
  • Author(s): O. Hisatomi and Y. Nakatani
  • Organizer: th Awaji International Workshop on “Electron Spin Science & Technology: Biological and Materials Science Oriented Applications” (AWEST 2016)
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] Quantitative analyses of the Photozipper/DNA complex formation (2016)
  • Author(s): Y. Nakatani and O. Hisatomi
  • Organizer: 4th Awaji International Workshop on “Electron Spin Science & Technology: Biological and Materials Science Oriented Applications” (AWEST 2016)
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] Regulation of a bZIP transcription factor by light (2015)
  • Author(s): Osamu Hisatomi
  • Organizer: 第40回日本比較内分泌学会・第37回日本比較生理生化学会 合同大会
  • Place of Presentation: JMSアステールプラザ、広島
  • Year and Date: 2015-12-11
• [Presentation] Evaluation of DNA-binding of a light-activatable bZIP module, Photozipper (2015)
  • Author(s): Osamu Hisatomi, Yuki Yabe, Yoichi Nakatani
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-13
• [Presentation] Molecular mechanism of Photozipper, a light-regulated bZIP module (2015)
  • Author(s): Yoichi Nakatani, Osamu Hisatomi
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-13
• [Presentation] Dark regeneration kinetics of site-directed mutants of bZIP module, Photozipper (2015)
  • Author(s): Yuki Yabe, Yoichi Nakatani, Osamu Hisatomi
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-13
• [Presentation] Analyses of a light-regulated bZIP protein, Photozipper, fused with YFP and mCherry (2015)
  • Author(s): Keigo Furuya, Osamu Hisatomi
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-13
• [Presentation] Characterization of an engineered photoactivatable bZIP module, Photozipper (2015)
  • Author(s): Osamu Hisatomi
  • Organizer: 3rd AWEST 2015
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2015-06-14
  • Int'l Joint Research / Invited
• [Presentation] A light-activatable bZIP module, Photozipper, induces the binding of fused proteins to the target sequence in a blue light-dependent manner (2015)
  • Author(s): Osamu Hisatomi and Keigo Furuya
  • Organizer: 3rd AWEST 2015
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2015-06-14
  • Int'l Joint Research
• [Presentation] Molecular mechanism of Photozipper, a light-regulated bZIP module (2015)
  • Author(s): Yoichi Nakatani, Osamu Hisatomi
  • Organizer: 3rd AWEST 2015
  • Place of Presentation: Awaji Island, Japan
  • Year and Date: 2015-06-14
  • Int'l Joint Research
• [Presentation] A photo-activated basic-leucine zipper module, opZL (2014)
  • Author(s): Hisatomi, O., Nakatani, Y. and Kai, Y.
  • Organizer: Interenational Conference of Retinal Proteins 2014
  • Place of Presentation: Nagahama, Shiga, Japan
  • Year and Date: 2014-10-05 – 2014-10-10
  • Invited
• [Presentation] Functional evaluation of the light-induced dimerizing module, Photodimerizer, fused with fluorescent proteins (2014)
  • Author(s): Osamu Hisatomi, Yoich Nakatani, Yuki Kai
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Molecular mechanism for dimerization of the light-regulated bZip module, Photodimeizer. (2014)
  • Author(s): Yoichi Nakatani, Osamu Hisatom
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Reaction Dynamics of Light Dependent Transcription Factor Aureochrome-1 (2014)
  • Author(s): Yuki Akiyama, Yusuke Nakasone, Osamu Hisatomi, Yoichi Nakatani, Masahide Terazima
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Photocontrolled DNA-binding of a bZIP protein. (2014)
  • Author(s): Hisatomi, O
  • Organizer: 2nd Awaji International Workshop on “Electron Spin Science & Technology: Biological and Materials Science Oriented Applications
  • Place of Presentation: Awaji Island, Hyogo, Japan
  • Year and Date: 2014-06-15 – 2014-06-17
  • Invited
• [Remarks] 久冨のページ
  • URL: http://gabriel.ess.sci.osaka-u.ac.jp/html/hisatomi/hisatomi-jp.html