Analysis of specific interactions among influenza A virus genomic RNAs

• Project/Area Number: 19K22529
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 49:Pathology, infection/immunology, and related fields
• 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): 2019-06-28 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2021: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: インフルエンザウイルス / ゲノムパッケージング / RNA

Outline of Research at the Start

インフルエンザウイルスのゲノムRNAは8分節に分かれている。各RNA分節はウイルス核タンパク質およびRNAポリメラーゼと結合し、RNP複合体として存在する。これまで、子孫ウイルス粒子が何本のRNA分節をどのように取り込むか、長年の謎だった。申請者は、各ゲノムRNA分節上にそれぞれ特有のパッケージングシグナル配列が必須であること、また、子孫ウイルス粒子内には8種類のRNP複合体が規則的な配置をとって取り込まれることを明らかにした。そこで本研究では、ゲノムパッケージング機構におけるウイルスゲノムRNAを介したRNP複合体間相互作用の立証とその分子基盤の解明を目的とする。

Outline of Final Research Achievements

Influenza A virus genome consists of eight-segmented, single-stranded, negative-sense viral RNAs (vRNAs). It has been shown that the eight vRNAs are selectively packaged into progeny virions through segment-specific packaging signals that are located in both the terminal coding regions and adjacent noncoding regions of each vRNA. However, the contributions of the packaging signals to the intersegment interactions are not understood. Here, using synthesized vRNAs containing the packaging signal sequences, we performed in vitro RNA binding assays and identified 15 intersegment interactions among eight vRNAs, most of which were mediated by the 3'- and 5'-terminal regions. Interestingly, all eight vRNAs interacted with multiple other vRNAs, in that some bound to different vRNAs through their respective 3'- and 5'-terminal regions. Our findings would be of use in future studies of in vivo vRNA-vRNA interactions during selective genome packaging.

Academic Significance and Societal Importance of the Research Achievements

本研究では、インフルエンザウイルスの8種類のゲノムRNAの間にパッケージングシグナルを介したRNA-RNA相互作用が存在することを明らかにした。本相互作用は選択的ゲノムパッケージングにおいて重要な役割を果たすと考えれらることから、新たな抗インフルエンザ薬の標的となると考えられる。

Research Products

• [Journal Article] Contribution of RNA-RNA Interactions Mediated by the Genome Packaging Signals for the Selective Genome Packaging of Influenza A Virus. (2022)
  • Author(s): Miyamoto S, Muramoto Y, Shindo K, Fujita-Fujiharu Y, Morikawa T, Tamura R, Gilmore JL, Nakano M, Noda T.
  • Journal Title: J Virol Volume: 96 Issue: 6
  • DOI: 10.1128/jvi.01641-21
  • Peer Reviewed / Open Access
• [Journal Article] Migration of Influenza Virus Nucleoprotein into the Nucleolus Is Essential for Ribonucleoprotein Complex Formation (2022)
  • Author(s): Sho Miyamoto, Masahiro Nakano, Takeshi Morikawa, Ai Hirabayashi, Ryoma Tamura, Yoko Fujita-Fujiharu, Nanami Hirose, Yukiko Muramoto, and Takeshi Noda
  • Journal Title: mBio Volume: 13 Issue: 1
  • DOI: 10.1128/mbio.03315-21
  • Peer Reviewed / Open Access
• [Journal Article] Ultrastructure of influenza virus ribonucleoprotein complexes during viral RNA synthesis (2021)
  • Author(s): Masahiro Nakano, Yukihiko Sugita, Noriyuki Kodera, Sho Miyamoto, Yukiko Muramoto, Matthias Wolf, and Takeshi Noda
  • Journal Title: Commun. Biol. Volume: 4 Issue: 1 Pages: 858-858
  • DOI: 10.1038/s42003-021-02388-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Selective Genome Packaging Mechanisms of Influenza A Viruses. (2021)
  • Author(s): Noda T
  • Journal Title: Cold Spring Harb Perspect Med. Volume: in press Pages: a038497-a038497
  • DOI: 10.1101/cshperspect.a038497
  • Peer Reviewed
• [Journal Article] Influenza A virus NS1 optimises virus infectivity by enhancing genome packaging in a dsRNA-binding dependent manner (2020)
  • Author(s): Sha Tim Wai、Weber Michaela、Kasumba Dacquin M.、Noda Takeshi、Nakano Masahiro、Kato Hiroki、Fujita Takashi
  • Journal Title: Virology Journal Volume: 17 Issue: 1 Pages: 107-107
  • DOI: 10.1186/s12985-020-01357-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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 Pages: 164-171
  • NAID: 40021927501
  • Peer Reviewed
• [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