Structural basis for nuclear transport mechanism by the nuclear transport receptor and improvement of a synchrotron beamline.

• Project Area: Structural basis of cell-signalling complexes mediating signal perception, transduction and responses
• Project/Area Number: 22121006
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: YAMASHITA Eiki 大阪大学, たんぱく質研究所, 助教 (00294132)
• Co-Investigator(Renkei-kenkyūsha): HIGASHIURA Akifumi 大阪大学, 蛋白質研究所, 特任助教 (90598129)
• Research Collaborator: YAMAZAWA Ryuji 大阪大学, 蛋白質研究所, 博士研究員 ; KOBAYASHI Takeshi 大阪大学, 大学院理学研究科 ; MATSUMOTO Ami 大阪大学, 大学院理学研究科
• Project Period (FY): 2010-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥65,390,000 (Direct Cost: ¥50,300,000、Indirect Cost: ¥15,090,000); Fiscal Year 2014: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000); Fiscal Year 2013: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000); Fiscal Year 2012: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000); Fiscal Year 2011: ¥28,470,000 (Direct Cost: ¥21,900,000、Indirect Cost: ¥6,570,000); Fiscal Year 2010: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 細胞構造生物 / 核-細胞質間輸送 / X線結晶構造解析 / 放射光 / 構造細胞生物 / 核細胞質間輸送因子 / 核外輸送因子 / RNA / Ran / tRNA

Outline of Final Research Achievements

Nucleocytoplasmic transport is essential for the function of eukaryotic cells. Nucleocytoplasmic transport of large molecules, including proteins and RNAs, require transport receptors. The transport receptors recognize cargo molecule and the receptor-cargo complexes are translocated through the nuclear pore complex. Here, we solved crystal structures of a transport receptor and transport receptor complex with the high accuracy measurement using a improved synchrotron beamline for the cell structural biological research.

Research Products

• [Journal Article] Structural and functional analysis of Hikeshi, a new nuclear transport receptor of Hsp70s. (2015)
  • Author(s): Song J, Kose S, Watanabe A, Son SY, Choi S, Hong H, Yamashita E, Park IY, Imamoto N, Lee SJ.
  • Journal Title: Acta Crystallogr D Biol Crystallogr. Volume: 71 Issue: 3 Pages: 473-483
  • DOI: 10.1107/s1399004714026881
  • Peer Reviewed
• [Journal Article] Structural basis for the selective nuclear import of the C2H2 zinc-finger protein Snail by importin β. (2014)
  • Author(s): Choi S., Yamashita E., Yasuhara N., Song J., Son S.Y., Won Y.H., Hong H.R., Shin Y.S., Sekimoto T., Park I.Y., Yoneda Y., Lee S.J.
  • Journal Title: Acta Crystallogr D Biol Crystallogr. Volume: 70 Issue: 4 Pages: 1050-1060
  • DOI: 10.1107/s1399004714000972
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray diffraction analysis of human importin β-Snail zinc finger domain complex. (2013)
  • Author(s): Choi S., Song J., Son S.Y., Park I.Y., Yamashita E., Lee S.J..
  • Journal Title: Acta Cryst. F Volume: 69 Issue: 6 Pages: 1049-1051
  • DOI: 10.1042/bst20120211
  • Peer Reviewed
• [Journal Article] Crystal structure of the C-terminal domain of Mu phage central spike and functions of bound calcium ion. (2013)
  • Author(s): Harada K
  • Journal Title: Biochim Biophys Acta., Volume: 1834 Issue: 1 Pages: 284-291
  • DOI: 10.1016/j.bbapap.2012.08.015
  • Peer Reviewed
• [Journal Article] Structural flexibility regulates phosphopeptide-binding activity of the tyrosine kinase binding domain of Cbl-c. (2012)
  • Author(s): Takesita K
  • Journal Title: J. Biochem. Volume: 152 Issue: 5 Pages: 487-495
  • DOI: 10.1093/jb/mvs085
  • Peer Reviewed
• [Journal Article] Structural insight into maintenance methylation by mouse DNA methyltransferase 1 (Dnmt1) (2011)
  • Author(s): Takeshita K., Yamashita E.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 108 Pages: 9055-9059
  • Peer Reviewed
• [Journal Article] The host-binding domain of the P2 phage tail spike reveals a trimeric iron-binding structure (2011)
  • Author(s): Yamashita E.
  • Journal Title: Ada Cryst Volume: F67 Pages: 837-841
  • Peer Reviewed
• [Journal Article] Selective nuclear export mechanism of small RNAs (2011)
  • Author(s): Lee S.J., Yamashita E.
  • Journal Title: Curre.Opin.Strcut.Biol. Volume: 21 Pages: 101-108
  • Peer Reviewed
• [Journal Article] Bovine cytochrome c oxidase structures enable O2 reduction with minimization of reactive oxygens and provide a proton-pumping gate (2010)
  • Author(s): Muramoto K., Yamashita E.
  • Journal Title: Proc.Natl.Acad.Sci.USA. Volume: 107 Pages: 7740-7745
  • Peer Reviewed
• [Presentation] SPring-8 BL44XU, Beamline Designed for a Structure Analysis of Large Biological Macromolecular Assemblies (2015)
  • Author(s): Akifumi Higashiura*, Eiki Yamashita*, Masato Yoshimura, Kazuya Hasegawa, Yukito Furukawa, Takashi Kumasaka, Go Ueno, Masaki Yamamoto, Tomitake Tsukihara, Atsushi Nakagawa
  • Organizer: The 12th International Conference on Synchrotron Radiation Instrumentation (SRI 2015)
  • Place of Presentation: New York City
  • Year and Date: 2015-07-06 – 2015-07-10
• [Presentation] 核外輸送におけるExportin-5の構造基盤 (2015)
  • Author(s): 山澤龍治，小林武，慈幸千真理，月原冨武，中川敦史，山下栄樹
  • Organizer: 第15回日本蛋白質科学会年会
  • Place of Presentation: あわぎんホール
  • Year and Date: 2015-06-24 – 2015-06-26
• [Presentation] SPring-8生体超分子複合体構造解析ビームライン（大阪大学蛋白質研究所）BL44XUの現状 (2015)
  • Author(s): 山下栄樹, 東浦彰史, 吉村政人, 長谷川和也, 熊坂崇，古川行人，大端通，上野剛，山本雅貴，吉川信也，月原冨武，中川敦史
  • Organizer: 第28回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 立命館大学びわこ・くさつキャンパス
  • Year and Date: 2015-01-10 – 2015-01-12
• [Presentation] IPR Beamline for Macromolecular Assemblies at SPring-8 (BL44XU) (2014)
  • Author(s): Eiki Yamashita, Akifumi Higashiura, Masato Yoshimura, Kazuya Hasegawa, Yukito Furukawa, Toru Ohata, Takashi Kumasaka, Go Ueno, Masaki Yamamoto, Atsushi Nakagawa
  • Organizer: 23rd Congress and General Assembly of the International Union of Crystallography (IUCr2014)
  • Place of Presentation: Montreal
  • Year and Date: 2014-08-05 – 2014-08-12
• [Presentation] SPring-8生体超分子複合体構造解析ビームライン（大阪大学蛋白質研究所）BL44XUの現状 (2014)
  • Author(s): 山下栄樹
  • Organizer: 第27回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 広島国際会議場（広島）
• [Presentation] SPring-8生体超分子複合体構造解析ビームライン（大阪大学蛋白質研究所）BL44XUの現状 (2013)
  • Author(s): 山下栄樹
  • Organizer: 第26回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 名古屋大学
  • Year and Date: 2013-01-12
• [Presentation] 異物排出タンパク質のX線結晶構造解析 (2013)
  • Author(s): 板倉勇太
  • Organizer: 第26回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 名古屋大学
  • Year and Date: 2013-01-12
• [Presentation] リガンドフリー型Exportin-5の構造解析 (2013)
  • Author(s): 山澤龍治
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 熊本大学（熊本）
• [Presentation] Exportin-5による RNAs の核外輸送メカニズムの解明 / The structural basis of RNAs nuclear export by Exportin-5. (2012)
  • Author(s): 山澤龍治
  • Organizer: 第12回 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2012-06-20
• [Presentation] SPring-8生体超分子複合体構造解析ビームライン(大阪大学蛋白質研究所)BL44XUの現状 (2012)
  • Author(s): 山下栄樹
  • Organizer: 第25回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 鳥栖市民文化会館・中央公民館
  • Year and Date: 2012-01-08
• [Presentation] SPring-8生体超分子複合体構造解析ビームライン(大阪大学蛋白質研究所)BL44XUの現状 (2011)
  • Author(s): 山下栄樹
  • Organizer: 第24回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: つくば国際会議場
  • Year and Date: 2011-01-10
• [Presentation] pre-microRNA核外輸送複合体のX線結晶構造解析 (2010)
  • Author(s): 山下栄樹
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートピアランド
  • Year and Date: 2010-12-10
• [Presentation] SPring-8阪大蛋白研ビームラインBL44XUの現状 (2010)
  • Author(s): 山下栄樹
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 大阪大学
  • Year and Date: 2010-12-03
• [Presentation] タンパク質合成を抑制する小さなRNAを輸送する仕組みの解明 (2010)
  • Author(s): 山下栄樹
  • Organizer: 第2回SPring-8合同コンファレンス
  • Place of Presentation: 東京ステーションコンファレンス(招待講演)
  • Year and Date: 2010-11-04