The novel molecular recognition mechanism of Ras by its structural polymorphism

• Project/Area Number: 25871145
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biophysics; Structural biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Nobuhisa Umeki 国立研究開発法人理化学研究所, 佐甲細胞情報研究室, 研究員 (70647502)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: Ras / RalGDS / 1分子計測 / 細胞内情報伝達 / 構造多型 / 一分子計測 / 低分子量Gタンパク質 / 低分子量G蛋白質

Outline of Final Research Achievements

To understand the mechanism of Ras-RalGDS-Ral signaling, translocation dynamics of RalGDS and its functional domains (RBD and REMCDC) to the plasma membranes of living cells were measured. Although the RBD played an important role in increasing the association rate constant between RalGDS and the plasma membrane, the REMCDC domain affected the dissociation rate constant from the membrane, which decreased after Ras activation. Thus, multiple regions and domains of both Ras and RalGDS work concertedly to regulate the Ras-RalGDS-Ral signaling. It is also suggested that the structural polymorphism of Ras is involved in interaction of Ras and RalGDS.

Research Products

• [Journal Article] Single-molecule fluorescence imaging of RalGDS on cell surfaces during signal transduction from Ras to Ral (2017)
  • Author(s): Yoshizawa R., Umeki N., Yanagawa M., Murata M., Sako Y.
  • Journal Title: Biophysics and Physicobiology Volume: -
  • NAID: 130007040995
  • Peer Reviewed / Open Access
• [Journal Article] Mutually exclusive cooperative binding of myosin and cofilin to actin filaments involves cooperative conformational changes of actin. (2016)
  • Author(s): Ngo KX, Umeki N, Kijima ST, Kodera N, Ueno H, Furutani-Umezu N, Nakajima J, Noguchi TQP, Nagasaki A, Tokuraku K, Uyeda TQP
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 35449-35449
  • DOI: 10.1038/srep35449
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cofilin-induced cooperative conformational changes of actin subunits revealed using cofilin-actin fusion protein (2016)
  • Author(s): Umeki N., Hirose K., Uyeda TQ
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 20406-20406
  • DOI: 10.1038/srep20406
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Interaction of a Novel Fluorescent GTP Analogue with the small G-protein K-Ras (2016)
  • 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
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The significance of membrane fluidity of feeder cell-derived substrates for maintenance of iPS cell stemness. (2015)
  • Author(s): Zhou Y, Mao H, Joddar B, Umeki N, Sako Y, Wada K, Nishioka C, Takahashi E, Wang Y, Ito Y.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 11386-11386
  • DOI: 10.1038/srep11386
  • Peer Reviewed / Open Access
• [Presentation] Elucidation of the EGF dependent localization mechanism of RalGDS molecule to plasma membrane using TIRF microscopy (2016)
  • Author(s): Yoshizawa R., Umeki N., Yanagawa M., Murata M., Sako Y.
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば国際会議場（茨城県つくば市）
  • Year and Date: 2016-11-25
• [Presentation] Photocontrol of small G-protein H-Ras multimer formation using caged compounds (2016)
  • Author(s): Iwata S., Hashimoto T., Umeki N., Sugimoto Y., Kondo K., Maruta S.
  • Organizer: American biophysical society 60th annual meeting
  • Place of Presentation: Los Angeles, USA
  • Year and Date: 2016-02-27
  • Int'l Joint Research
• [Presentation] Ras-RalGDS相互作用の1分子蛍光イメージング (2015)
  • Author(s): 梅木伸久、吉澤亮、中村由樹、稲葉岳彦、前田亮、柳川正隆、佐甲靖志
  • Organizer: 第88回日本生化学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
  • Int'l Joint Research
• [Presentation] ケイジド化合物導入による低分子量Gタンパク質H-Rasの多量体形成の光制御 (2015)
  • Author(s): 岩田聖悟、橋本貴志、梅木伸久、近藤和典、丸田晋策
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢
  • Year and Date: 2015-09-13
  • Int'l Joint Research
• [Presentation] Photo-regulation of the interaction between ras and ralgds using GTP analogues composed of photochromic molecules (2015)
  • Author(s): Masuhara K, Iwata S, Umeki N, Maruta S
  • Organizer: American bio-physics 59th annual meeting
  • Place of Presentation: Baltimore USA
  • Year and Date: 2015-02-07 – 2015-02-11
• [Presentation] Photo-regulation of small G protein normal and oncogenic K-ras using Photochromic molecules. (2015)
  • Author(s): Iwata S, Masuhara K, Umeki N, Kondo K, Maruta S
  • Organizer: American bio-physics 59th annual meeting
  • Place of Presentation: Baltimore USA
  • Year and Date: 2015-02-07 – 2015-02-11
• [Presentation] photo-regulation of small G-pritein K-ras using photochromic molecules (2014)
  • Author(s): Iwata S, Masuhara K, Umeki N, Kondo K, Maruta S
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-09-25 – 2014-09-27