Molecular and structural basis for transcription and replication mechanism of influenza virus genome

• Project/Area Number: 17H04082
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: Kyoto University
• Principal Investigator: NODA TAKESHI 京都大学, ウイルス・再生医科学研究所, 教授 (00422410)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2017: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: インフルエンザウイルス / RNP / 転写 / 複製 / 転写・複製 / ゲノム

Outline of Final Research Achievements

Influenza virus genomic RNA (vRNA) constitutes a double-helical ribonucleoprotein complex (RNP complex) together with viral nucleoproteins and a viral RNA polymerase, which is responsible for the transcription and replication of the vRNA. However, it remains uncertain whether the double-helical structure of the RNP complex shows confomational changes during transcription and replication. Here by using high speed atomic force microscopy and cryoelectron microscopy, we investigated the ultrastructural changes of the helical RNPs during in vitro RNA synthesis reaction. We found that there are double-helical RNPs sassociated with newly synthesized RNA, but some RNPs associated with newly synthesized RNA are deformed.

Academic Significance and Societal Importance of the Research Achievements

インフルエンザウイルスのゲノムRNAはウイルス核タンパク質NPやポリメラーゼと共にRNP複合体を形成する。RNP複合体はウイルスRNAの転写および複製を担う分子装置として機能する。本研究ではRNP複合体が転写および複製時の構造に着目し、高速原子間力顕微鏡およびクライオ電子顕微鏡を用いてその微細構造を解析した。その結果、RNA合成時にRNP複合体は2つの形態をとることが明らかになった。これらの成果は転写複製を担う分子装置のRNA合成機構を理解する上で重要な知見となる。

Research Products

• [Journal Article] G Protein Pathway Suppressor 1 Promotes Influenza Virus Polymerase Activity by Activating the NF-κB Signaling Pathway. (2020)
  • Author(s): Kuwahara T, Yamayoshi S, Noda T, Kawaoka Y.
  • Journal Title: mBio Volume: 10 Issue: 6
  • DOI: 10.1128/mbio.02867-19
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In vitro vRNA-vRNA interactions in the H1N1 influenza A virus genome. (2020)
  • Author(s): Miyamoto S, Noda T.
  • Journal Title: Microbiol Immunol Volume: 64 Issue: 3 Pages: 202-209
  • DOI: 10.1111/1348-0421.12766
  • Peer Reviewed
• [Journal Article] N4BP1 restricts HIV-1 and its inactivation by MALT1 promotes viral reactivation. (2019)
  • Author(s): Yamasoba D, Sato K, Ichinose T, Imamura T, Koepke L, Joas S, Reith E, Hotter D, Misawa N, Akaki K, Uehata T, Mino T, Miyamoto S, Noda T, Yamashita A, Standley DM, Kirchhoff F, Sauter D, Koyanagi Y, Takeuchi O.
  • Journal Title: Nat Microbiol Volume: 4 Issue: 9 Pages: 1532-1544
  • DOI: 10.1038/s41564-019-0460-3
  • NAID: 120006731123
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of a purely clonal defective interfering influenza virus. (2019)
  • Author(s): Yamagata Y, Muramoto Y, Miyamoto S, Shindo K, Nakano M, Noda T.
  • Journal Title: Microbiol Immunol. Volume: 63 Issue: 5 Pages: 164-171
  • DOI: 10.1111/1348-0421.12681
  • NAID: 40021927501
  • Peer Reviewed / Open Access
• [Journal Article] Local structural changes of the influenza A virus ribonucleoprotein complex by single mutations in the specific residues involved in efficient genome packaging. (2019)
  • Author(s): Takizawa N, Ogura Y, Fujita Y, Noda T, Shigematsu H, Hayashi T, Kurokawa K
  • Journal Title: Virology Volume: 531 Pages: 126-140
  • DOI: 10.1016/j.virol.2019.03.004
  • Peer Reviewed / Open Access
• [Journal Article] A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages. (2018)
  • Author(s): Bedi S, Noda T, Kawaoka Y, Ono A.
  • Journal Title: MBio Volume: 9 Issue: 5
  • DOI: 10.1128/mbio.01916-18
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Influenza A virus nucleoprotein is acetylated by histone acetyltransferases, PCAF and GCN5 (2018)
  • Author(s): Hatakeyama D、Shoji M、Yamayoshi S、Yoh R、Ohmi N、Takenaka S、Saitoh A、Arakaki Y、Masuda A、Komatsu T、Nagano R、Nakano M、Noda T、Kawaoka Y、Kuzuhara T
  • Journal Title: Journal of Biological Chemistry Volume: in press Issue: 19 Pages: 7126-7138
  • DOI: 10.1074/jbc.ra117.001683
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Lung-derived exosomal miR-483-3p regulates the innate immune response to influenza virus infection. (2018)
  • Author(s): Maemura T, Fukuyama S, Sugita Y, Lopes TJS, Nakao T, Noda T, Kawaoka Y.
  • Journal Title: J Infect Dis Volume: 217 Issue: 9 Pages: 1372-1382
  • DOI: 10.1093/infdis/jiy035
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Influenza C and D Viruses Package Eight Organized Ribonucleoprotein Complexes (2018)
  • Author(s): Nakatsu Sumiho、Murakami Shin、Shindo Keiko、Horimoto Taisuke、Sagara Hiroshi、Noda Takeshi、Kawaoka Yoshihiro
  • Journal Title: Journal of Virology Volume: 92 Issue: 6
  • DOI: 10.1128/jvi.02084-17
  • Peer Reviewed
• [Journal Article] Importance of the 1+7 configuration of ribonucleoprotein complexes for influenza A virus genome packaging. (2018)
  • Author(s): Noda T, Murakami S, Nakatsu S, Imai H, Muramoto Y, Shindo K, Sagara H, Kawaoka Y.
  • Journal Title: Nat Commun. Volume: 9 Issue: 1 Pages: 59-59
  • DOI: 10.1038/s41467-017-02517-w
  • NAID: 120006376026
  • Peer Reviewed / Open Access / Int'l Joint Research