Identification and characterization of novel influenza A virus proteins

• Project/Area Number: 26460549
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Virology
• Research Institution: The University of Tokyo
• Principal Investigator: Yamayoshi Seiya 東京大学, 医科学研究所, 特任准教授 (50529534)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: インフルエンザウイルス / PB2分節 / スプライス / 新規ウイルス蛋白質 / スプライシング / PB2

Outline of Final Research Achievements

Over the past two decades, several novel influenza viral proteins have been identified that modulate viral infections in vitro and/or in vivo. We used RT-PCR targeting viral mRNAs transcribed from the PB2 segment to look for novel viral proteins encoded by spliced mRNAs. We identified a new viral protein, termed PB2-S1, encoded by a novel spliced mRNA. PB2-S1 was detected in virus-infected cells and in cells transfected with a plasmid encoding PB2. PB2-S1 localized to mitochondria, inhibited the RIG-I-dependent interferon signaling pathway, and interfered with viral polymerase activity depending on its PB1 binding capability. The nucleotide sequences around the splicing donor and acceptor sites for PB2-S1 were highly conserved among human pre-2009 H1N1 viruses but not among human H1N1pdm and H3N2 viruses. PB2-S1-deficient viruses, however, showed similar growth kinetics in MDCK cells and virulence in mice to those of wild-type virus.

Research Products

• [Journal Article] Evaluation of seasonal influenza vaccinese for H1N1pdm09 and type B viruses based on a replication-incompetent PB2-KO virus. (2017)
  • Author(s): Ui H, Yamayoshi S, Uraki R, Kiso M, Oishi K, Murakami S, Mimori S, Kawaoka Y.
  • Journal Title: Vaccine Volume: 4;35(15) Issue: 15 Pages: 1892-1897
  • DOI: 10.1016/j.vaccine.2017.02.041
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The host protein CLUH participates in the subnuclear transport of influenza virus ribonucleoprotein complexes (2016)
  • Author(s): Ando T, Yamayoshi S, Tomita Y, Watanabe T, Watanabe S, Kawaoka Y
  • Journal Title: Nature Microbiology Volume: 1 Issue: 8 Pages: 16062-16062
  • DOI: 10.1038/nmicrobiol.2016.62
  • Peer Reviewed / Open Access
• [Journal Article] Risk assessment of recent Egyptian H5N1 influenza viruses (2016)
  • Author(s): Arafa AS, Yamada S, Imai M, Watanabe T et al
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 38388-38388
  • DOI: 10.1038/srep38388
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The Microminipig as an Animal Model for Influenza A Virus Infection (2016)
  • Author(s): Iwatsuki-Horimoto K, Nakajima N, Shibata M, Takahashi K, Sato Y, Kiso M, Yamayoshi S, Ito M, Enya S, Otake M, Kangawa A, da Silva Lopes TJ, Ito H, Hasegawa H, Kawaoka Y.
  • Journal Title: Journal of Virology Volume: 91 Issue: 2 Pages: 1-9
  • DOI: 10.1128/jvi.01716-16
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A Broadly Reactive Human Anti-hemagglutinin Stem Monoclonal Antibody That Inhibits Influenza A Virus Particle Release. (2016)
  • Author(s): Yamayoshi S, Uraki R, Ito M, Kiso M, Nakatsu S, Yasuhara A, Oishi K, Sasaki T, Ikuta K,Kawaoka Y.
  • Journal Title: EBioMedicine Volume: 17 Pages: 182-191
  • DOI: 10.1016/j.ebiom.2017.03.007
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Viral Protein Expressed from the PB2 Segment of Influenza A Virus. (2016)
  • Author(s): Yamayoshi S, Watanabe M, Goto H, Kawaoka Y.
  • Journal Title: Journal of Virology Volume: 90 Issue: 1 Pages: 444-456
  • DOI: 10.1128/jvi.02175-15
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Amino acids substitutions in the PB2 protein of H7N9 influenza A viruses are important for virulence in mammalian hosts. (2015)
  • Author(s): Yamayoshi S, Fukuyama S, Yamada S, Zhao D, Murakami S, Uraki R, Watanabe T, Tomita Y, Neumann G, Kawaoka Y.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 8039-8039
  • DOI: 10.1038/srep08039
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Ａ ｎｏｖｅｌ ｆｕｎｃｔｉｏｎ ｓｉｔｅ ｉｎ ｔｈｅ ＰＢ２ ｓｕｂｕｎｉｔ ｏｆ ｉｎｆｌｕｅｎｚａ Ａ ｖｉｒｕｓ ｅｓｓｅｎｔｉａｌ ｆｏｒ ａｃｅｔｙｌ－ＣｏＡ ｉｎｔｅｒａｃｔｉｏｎ， ＲＮＡ ｐｏｌｙｍｅｒａｓｅ ａｃｔｉｖｉｔｙ， ａｎｄ ｖｉｒａｌ ｒｅｐｌｉｃａｔｉｏｎ． (2014)
  • Author(s): Ｈａｔｋｅｙａｍａ Ｄ, Shoji M, Yamayoshi S, Hirota T, Nagae M, Yanagisawa S, Nakano M, Ohmi N, Noda T, Kawaoka Y, Kuzuhara T.
  • Journal Title: Ｊ Ｂｉｏｌ Ｃｈｅｍ Volume: 289 Issue: 36 Pages: 24980-94
  • DOI: 10.1074/jbc.m114.559708
  • Peer Reviewed / Open Access
• [Journal Article] Virulence-Affecting Amino Acid Changes in the PA Protein of H7N9 Influenza A Viruses. (2014)
  • Author(s): Yamayoshi S, Yamada S, Fukuyama S, Murakami S, Zhao D, Uraki R, Watanabe T, Tomita Y, Macken C, Neumann G, Kawaoka Y
  • Journal Title: Journal of Virology Volume: 88(6) Issue: 6 Pages: 3127-34
  • DOI: 10.1128/jvi.03155-13
  • Peer Reviewed
• [Presentation] A broadly reactive human anti-hemagglutinin stem monoclonal antibody that inhibits influenza A virus particle release (2017)
  • Author(s): Yamayoshi S, Uraki R, Ito M, Kiso M, Nakatsu S, Yasuhara A, Oishi K, Sasaki T, Ikuta K, Kawaoka Y.
  • Organizer: The 6th China‐Japan Bilateral Symposium on All Influenza virus
  • Place of Presentation: 中国・北京
  • Year and Date: 2017-03-14
  • Int'l Joint Research / Invited
• [Presentation] A broadly reactive human anti-HA monoclonal antibody that inhibits influenza A virus particle release (2016)
  • Author(s): S. Yamayoshi, R. Uraki, M. Ito, M. Kiso, S. Nakatsu, K. Oishi, A. Yasuhara, T. Sasaki, K. Ikuta, Y. Kawaoka
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2016-10-23
• [Presentation] Acetylation of NP of influenza virus affects viral transcription activities (2016)
  • Author(s): D. Hatakeyama, M. Shoji, S. Yamaysohi, R. Yho, N. Ohmi, S. Takenaka, Y. Arakaki, A. Masuda, T. Komatsu, A. Saitoh, M. Nakano, T. Noda, Y. Kawaoka, T. Kuzuhara
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2016-10-23
• [Presentation] Analysis of the regulatory mechanisms of the subnuclear transport of influenza progeny ribonucleoprotein complexes (2016)
  • Author(s): T. Ando, S. Yamaysohi, Y. Tomita, S. Watanabe, T. Watanabe, Y. Kawaoka
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2016-10-23
• [Presentation] Toward the identification of host proteins involved in the shutoff activity of influenza A virus PA-X (2016)
  • Author(s): K. Oishi, S. Yamayoshi, Y. Kawaoka
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2016-10-23
• [Presentation] Characterization of the antigenic properties of influenza A(H1N1)pdm09 virus (2016)
  • Author(s): A. Yasuhara, S. Yamayoshi, M. Ito, R. Uraki, S. Nakatsu, K. Oishi, P. Soni, T. Takenaga, C. Kawakami, E. Takashita, T. Sasaki, K. Ikuta, S. Yamada, Y. Kawaoka
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2016-10-23
• [Presentation] Involvement of CLUH in the subnuclear transport of influenza progeny ribonucleoprotein complexes (2016)
  • Author(s): T. Ando, S. Yamayoshi, S. Watanabe, T. Watanabe, Y. Kawaoka
  • Organizer: The Options IX for the Control of Influenza
  • Place of Presentation: アメリカ・シカゴ
  • Year and Date: 2016-08-24
  • Int'l Joint Research
• [Presentation] A broadly reactive human anti-HA monoclonal antibody that inhibits influenza A virus particle release (2016)
  • Author(s): S. Yamayoshi, R. Uraki, M. Ito, M. Kiso, S. Nakatsu, K. Oishi, A. Yasuhara, T. Sasaki, K. Ikuta, Y. Kawaoka
  • Organizer: The Options IX for the Control of Influenza
  • Place of Presentation: アメリカ・シカゴ
  • Year and Date: 2016-08-24
  • Int'l Joint Research
• [Presentation] Toward the identification of host proteins involved in the shutoff activity of influenza A virus PA-X (2016)
  • Author(s): K. Oishi, S. Yamayoshi, Y. Kawaoka
  • Organizer: The Options IX for the Control of Influenza
  • Place of Presentation: アメリカ・シカゴ
  • Year and Date: 2016-08-24
  • Int'l Joint Research
• [Presentation] Characterization of the antigenic properties of influenza A(H1N1)pdm09 virus (2016)
  • Author(s): A. Yasuhara, S. Yamayoshi, M. Ito, R. Uraki, S. Nakatsu, K. Oishi, P. Soni, T. Takenaga, T. Sasaki, K. Ikuta, S. Yamada, Y. Kawaoka
  • Organizer: The Options IX for the Control of Influenza
  • Place of Presentation: アメリカ・シカゴ
  • Year and Date: 2016-08-24
  • Int'l Joint Research
• [Presentation] Identification of a novel viral protein expressed from the PB2 segment of influenza A virus (2015)
  • Author(s): S. Yamayoshi, M. Watanabe, H. Goto, M. Hatta, Y. Kawaoka
  • Organizer: 16th Negative Strand Virus Meeting 2015
  • Place of Presentation: イタリア・シエナ
  • Year and Date: 2015-06-14
  • Int'l Joint Research
• [Presentation] A 型インフルンザウイルスが発現するPA_X の蛋白質発現抑制に重要な領域の同定 (2014)
  • Author(s): 大石康平、山吉誠也、河岡義裕
  • Organizer: 第62回日本ウイルス学会学術集会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-10
• [Presentation] H7N9鳥インフルエンザウイルスのPB2蛋白質の627Kおよび701Dは哺乳類に対する高病原性に寄与する (2014)
  • Author(s): 山吉誠也、福山聡、山田晋弥、Dongming Zhao、村上晋、浦木隆太、渡辺登喜子、百瀬(冨田) 有里子、Gabriele Neumann、河岡義裕
  • Organizer: 第157回 日本獣医学会学術集会
  • Place of Presentation: 北海道大学（北海道札幌市）
  • Year and Date: 2014-09-10
• [Presentation] Identification of a novel viral protein expressed from the PB2 segment of influenza A virus (2014)
  • Author(s): S. Yamayoshi, M. Watanabe, R. Uraki, H. Goto, and Y. Kawaoka
  • Organizer: 16th International Congress of Virology
  • Place of Presentation: モントリオール（カナダ）
  • Year and Date: 2014-07-28
• [Presentation] ウイルス増殖能に対するPB2 サブユニットにおける新規機能部位VRG 配列の役割 (2014)
  • Author(s): 畠山 大、庄司正樹、山吉誠也、廣田丈典、長江萌菜美、柳澤 伸、河岡義裕、葛原 隆
  • Organizer: 第28回インフルエンザ研究者交流の会シンポジウム
  • Place of Presentation: 鳥取市総合福祉センター（鳥取県鳥取市）
  • Year and Date: 2014-07-04