Structural basis for an essential subunit interaction in influenza virus RNA polymerase PB1-PB2

• Project/Area Number: 21370046
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Yokohama City University
• Principal Investigator: PARK Sam-yong 横浜市立大学, 生命ナノシステム科学研究科, 教授 (20291932)
• Co-Investigator(Kenkyū-buntansha): FUKUNISHI Yoshihumi 産業技術総合研究所, バイオメディシナル情報研究センター, 主任研究員 (60357895)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000); Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2010: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000); Fiscal Year 2009: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
• Keywords: インフルエンザ / RNAポリメラーゼ / 構造解析 / 創薬 / 複合体 / 構造我解析

Research Abstract

Influenza virus RNA-dependent RNA polymerase is a multi-functional heterotrimer, which uses a "cap-snatching" mechanism to produce viral mRNA. Host cell mRNA is cleaved to yield a cap-bearing oligonucleotide which can be extended using viral genomic RNA as a template. The cap binding and endonuclease activities are only activated once viral genomic RNA is bound. This requires signaling from the RNA-binding PB1 subunit to the cap-binding PB2 subunit, and the interface between these two subunits is essential for polymerase activity. We have defined this interaction surface by protein crystallography, and tested the effects of mutating contact residues on the function of the holo-enzyme. This novel interface is surprisingly small, yet it plays a crucial role in regulating the 250 kDa polymerase, and is completely conserved among avian and human influenza.

Research Products

• [Journal Article] Oxygen binding and redox properties of the heme in soluble guanylate cyclase : Implications for the mechanism of ligand discrimination (2011)
  • Author(s): Makino R, Park SY, Obayashi E, Iizuka T, Hori H, Shiro Y.
  • Journal Title: J Biol Chem Volume: 286(18) Pages: 15678-87
• [Journal Article] Crystal structure of the human centromeric nucleosome containing CENP-A (2011)
  • Author(s): Tachiwana H, Kagawa W, Shiga T, Osakabe A, Miya Y, Saito K, Hayashi-Takanaka Y, Oda T, Sato M, Park SY, Kimura H, Kurumizaka H.
  • Journal Title: Nature Volume: 476(7359) Pages: 232-5
• [Journal Article] Structures and oxygen affinities of crystalline human hemoglobin C({beta} 6 Glu-> Lys) in the R and R2 quaternary structures (2011)
  • Author(s): Shibayama N, Sugiyama K, Park SY
  • Journal Title: J Biol Chem Volume: 286(38) Pages: 33661-8
• [Journal Article] Crystal Structures of Penicillin-Binding Proteins 4 and 5 from Haemophilus influenzae (2010)
  • Author(s): Kawai F, Clarke TB, Roper DI, Han GJ, Hwang KY, Unzai S, Obayashi E, Park SY, Tame JR
  • Journal Title: J Mol Biol Volume: 396(3) Pages: 634-45
• [Journal Article] Direct determination of protonation states of histidine residues in a 2 A neutron structure of deoxy-human normal adult hemoglobin and implications for the Bohr effect (2010)
  • Author(s): Kovalevsky AY, Chatake T, Shibayama N, Park SY, Ishikawa T, Mustyakimov M, Fisher Z, Langan P, Morimoto Y.
  • Journal Title: J Mol Biol Volume: 398(2) Pages: 276-91
• [Journal Article] Autotransporter passenger proteins : virulence factors with common structural themes (2010)
  • Author(s): Nishimura K, Tajima N, Yoon YH, Park SY, Tame JR
  • Journal Title: J Mol Med Volume: 88(5) Pages: 451-8
• [Journal Article] A novel intein-like autoproteolytic mechanism in autotransporter proteins (2010)
  • Author(s): Tajima N, Kawai F, Park SY, Tame JR
  • Journal Title: J Mol Biol Volume: 402(4) Pages: 645-56
• [Journal Article] Role of domains within the autotransporter Hbp/Tsh (2010)
  • Author(s): Nishimura K, Yoon YH, Kurihara A, Unzai S, Luirink J, Park SY, Tame JR
  • Journal Title: Acta Crystallogr D Biol Crystallogr Volume: 66(12) Pages: 1295-300
• [Journal Article] 新規抗インフルエンザウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Journal Title: 生物物理 Volume: Vol.50, No.2 Pages: 74-79
  • NAID: 10025976244
• [Journal Article] タンパク質発現プロトコール (2010)
  • Author(s): 浅賀正充、朴三用
  • Journal Title: 実験医学別冊(羊土社) Pages: 94-111
• [Journal Article] インフルエザウスRNAポリメラーゼの構造解析と創薬研究 (2010)
  • Author(s): 杉山佳奈子、尾林栄治、吉田尚史、朴三用
  • Journal Title: 日本結晶学会 Volume: 52 Pages: 271-278
• [Journal Article] インフルエンザウイルスRNAポリメラーゼの構造解析と創薬研究 (2010)
  • Author(s): 杉山佳奈子、尾林栄治、吉田尚史、朴三用
  • Journal Title: 日本結晶学会 Volume: 52 Pages: 271-278
  • NAID: 10027673105
  • Peer Reviewed
• [Journal Article] Structural insight into the essential PB1-PB2 subunit contact of the influenza virus RNA polymerase (2009)
  • Author(s): Sugiyama K, Obayashi E, Kawaguchi A, Suzuki Y, Tame JR, Nagata K, Park SY.
  • Journal Title: EMBO J Volume: 28(12) Pages: 1803-11
• [Journal Article] Structural Studies of a Bacterial Condensin Complex Reveal ATP-Dependent Disruption of Intersubunit Interactions (2009)
  • Author(s): Woo JS, Lim JH, Shin HC, Suh MK, Ku B, Lee KH, Joo K, Robinson H, Lee J, Park SY, Ha NC, Oh BH.
  • Journal Title: Cell Volume: 136(1) Pages: 85-96
• [Journal Article] The nature of the TRAP-Anti-TRAP complex (2009)
  • Author(s): Watanabe M, Heddle JG, Kikuchi K, Unzai S, Akashi S, Park SY, Tame JR
  • Journal Title: Proc Natl Acad Sci U S A Volume: 106(7) Pages: 2176-81
• [Journal Article] Visualizing breathing motion of internal cavities in concert with ligand migration in myoglobin (2009)
  • Author(s): Tomita A, Sato T, Ichiyanagi K, Nozawa S, Ichikawa H, Chollet M, Kawai F, Park SY, Tsuduki T, Yamato T, Koshihara SY, Adachi SI
  • Journal Title: Proc Natl Acad Sci U S A Volume: 106(8) Pages: 2612-6
• [Journal Article] Mutational Analysis of the Substrate Specificity of Escherichia coli Penicillin Binding Protein 4(PBP4) (2009)
  • Author(s): Clarke T, Kawai F, Park SY, Tame J, Dowson C, Roper D.
  • Journal Title: Biochemistry Volume: 48(12) Pages: 2675-83
• [Journal Article] RNA and Protein Complexes of trp RNA-Binding Attenuation Protein Characterized by Mass Spectrometry (2009)
  • Author(s): Akashi S, Watanabe M, Heddle JG, Unzai S, Park SY, Tame JR
  • Journal Title: Anal Chem Volume: 81(6) Pages: 2218-26
• [Journal Article] 抗インフルエンザ薬の開発研究 (2009)
  • Author(s): 朴三用
  • Journal Title: Bipphiliaビオフィリア(アドスリ社) Volume: 5(4) Pages: 25-29
• [Journal Article] Structural insight into the essential PB1-PB2 subunit contact of the influenza virus RNA polymerase (2009)
  • Author(s): Sugiyama K, Obayashi E, Kawaguchi A, Suzuki Y, Tame JR, Nagata K, Park SY
  • Journal Title: EMBO J. 28(12) Pages: 286-92
  • Peer Reviewed
• [Presentation] Structure of PB1-PB2 subunit interface of inflenza A virus RNA polymerase (2011)
  • Author(s): Sugiyama K, Obayashi E, Kawaguchi A, Suzuki Y, Tame JR, Nagata K, Park SY
  • Organizer: IUCr
  • Place of Presentation: Palacio Municipal de Congresos. Madrid, Spain
• [Presentation] The structural basis for the essential PA-PB1 subunit interaction in influenza RNA polymerase (2011)
  • Author(s): Yoshida H, Obayashi E, Kawai F, Shibayama N, Kawaguchi A, Nagata K, Tame JR, Park SY
  • Organizer: IUCr
  • Place of Presentation: Palacio Municipal de Congresos. Madrid, Spain
• [Presentation] Structural Studies of the Influenza RNA-polymerase for Novel Drug Design (2011)
  • Author(s): Sam-Yong Park
  • Organizer: The 9^<th> International Symposium for Future Drug Design and Medical Care
  • Place of Presentation: Hokkaido University
• [Presentation] Structural Studies of the Influenza RNA-polymerase for Novel Drug Design (2011)
  • Author(s): Park SY
  • Organizer: The 9^<th> International Symposium for Future Drug Design and Medical Care
  • Place of Presentation: Hokkaido University(招待講演)
• [Presentation] 新規抗ウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 平成22年度KASTフォーラム1ドイツバイオ交流フォーラム
  • Place of Presentation: 川崎
  • Year and Date: 2010-07-02
• [Presentation] 新規抗インフルエンサウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 第27回PFシンポジウム
  • Place of Presentation: 筑波
• [Presentation] インフルエンザRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 第47回日本蛋白質科学会
  • Place of Presentation: 北海道大学
• [Presentation] インフルエンザウイルスのRNAポリメラーゼサブユニットPB1-PB2サブユニト間相互作用の解明 (2010)
  • Author(s): 杉山佳奈子、尾林栄治、川口敦史、鈴木由佳理、テイムジェレミー、永田恭介、朴三用
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌
• [Presentation] インフルエンザウイルスのRNAポリメラーゼサブユニットPA-PB1サブユニト間相互作用の解明 (2010)
  • Author(s): 吉田尚史、尾林栄治、河合文啓、川口敦史、永田恭介、テイムジェレミー、朴三用
  • Organizer: 第10回日本蛋白質科学会年会
  • Place of Presentation: 札幌
• [Presentation] 新規抗インフルエンザウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 第34回阿蘇シンポジウム
  • Place of Presentation: 阿蘇
• [Presentation] Structural Insight into the Subunit Contact of the Influenza Virus RNA Polymerase (2010)
  • Author(s): Sam-Yong Park
  • Organizer: 2^<nd> International Symposium on Drug Discovery and Design by NMR
  • Place of Presentation: Yokohama City University
• [Presentation] 新規抗ウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 大阪大学蛋白質研究所セミナー
• [Presentation] インフルエンザRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 第47回日本蛋白質科学会
  • Place of Presentation: 北海道大学(招待講演)
• [Presentation] インフルエンザウイルスのRNAポリメラーゼサブユニットPB1-PB2サブユニット間相互作用の解明 (2010)
  • Author(s): 杉山佳奈子、尾林栄治、川口敦史、鈴木由佳理、テイム ジェレミー、永田恭介、朴三用
  • Organizer: 第47回日本蛋白質科学会
  • Place of Presentation: 北海道大学
• [Presentation] インフルエンザウイルスのRNAポリメラーゼサブユニットPA-PB1サブユニット間相互作用の解明 (2010)
  • Author(s): 吉田尚史、尾林栄治、河合文啓、川口敦史、永田恭介、テイム ジェレミー、朴三用
  • Organizer: 第47回日本蛋白質科学会
  • Place of Presentation: 北海道大学
• [Presentation] 新規抗インフルエンザウイルス薬の開発基盤となるRNAポリメラーゼの構造解析 (2010)
  • Author(s): 朴三用
  • Organizer: 第34回 阿蘇シンポジウム
  • Place of Presentation: 阿蘇(招待講演)
• [Presentation] Structural Insight into the Subunit Contact of the Influenza Virus RNA Polymerase (2010)
  • Author(s): 朴三用
  • Organizer: 2^<nd> International Symposium on Drug Discovery and Design by NMR
  • Place of Presentation: Yokohama(招待講演)
• [Presentation] Structural Insight in to the Essential PA-PB1 Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): Sam-Yong Park
  • Organizer: VII European Symposium of the Protein Society
  • Place of Presentation: Zurich, Switzerland
• [Presentation] Structural Insight in to the Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): Sam-Yong Park
  • Organizer: America Crystallographic Association
  • Place of Presentation: Toronto Canada
• [Presentation] Structural Insight into the Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): Sam-Yong Park
  • Organizer: 9th KIAS-SUN Structure and Function
  • Place of Presentation: Seoul National University, Seoul, Korea
• [Presentation] Structural Insight into the Essential Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): E. Obayashi, K. Sugiyama, H. Yoshida, F. Kawai, A. Kawaguchi, Y. Suzuki, J. Tame, K. Nagata, SY Park
  • Organizer: 第47回日本生物物理学会
  • Place of Presentation: 徳島文理大学
• [Presentation] Structure and function of Root effect fish hemoglobin (2009)
  • Author(s): S. Unzai, K. Imai, T. Yokoyama, SY Park, K. Nagai, J. Tame
  • Organizer: 第47回日本生物物理学会
  • Place of Presentation: 徳島文理大学
• [Presentation] Structural Insight into the Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): Sam-Yong Park
  • Organizer: The 9th R. O. C-Japan Joint Seminar on Crystallography
  • Place of Presentation: Osaka University
• [Presentation] Structural Insight into the Subunit Contact of the Influenza Virus RNA Polymerase (2009)
  • Author(s): Sam-Yong Park
  • Organizer: America Crystallographic Association
  • Place of Presentation: Toronto Canada
• [Book] トピックス:万能インフルエンザ治療薬に道筋 (2009)
  • Author(s): 朴三用
  • Total Pages: 117
  • Publisher: メディカル
• [Book] メディカル・トピックス:万能インフルエンザ治療薬に道筋 (2009)
  • Author(s): 朴三用
  • Total Pages: 1
  • Publisher: Newton 8月号
• [Remarks]
  • URL: http://www.tsurumi.yokohama-cu.ac.jp/pdl/index.html
• [Patent(Industrial Property Rights)] インフルエンザウイルス由来のNAポリメラーゼ発現系構築と結晶化及び、抗インフルエンザ薬のスクリーニング方法 (2009)
  • Inventor(s): 朴三用、尾林栄治、永田恭介、川口敦史
  • Industrial Property Rights Holder: 公立大学法人横浜市立大学、国立大学法人筑波大学
  • Filing Date: 2009-07-02
  • Overseas