Construction of baculovirus-independent highly efficient protein expression system in insect cells

• Project/Area Number: 17K08162
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Insect science
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Nagamine Toshihiro 国立研究開発法人理化学研究所, 開拓研究本部, 専任研究員 (90237553)
• Project Period (FY): 2017-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: バキュロウイルス / 昆虫細胞 / タンパク質合成系 / 昆虫

Outline of Final Research Achievements

This study was initiated to construct a baculovirus-independent highly efficient protein expression system in insect cells using a virus-induced host cell shut-off mechanism. Although we could not construct the expression system, we found structural changes of IE1, a protein that induces liquid-liquid phase separation (LLPS), which is thought to be related to the shut-off mechanism, with the infection stage, and found that IE1 is an important conformational switch for LLPS formation.

Academic Significance and Societal Importance of the Research Achievements

バキュロウイルスの転写はvirogenic stroma（VS）と呼ばれるウイルスが誘導する核内構造体、すなわちLLPS内で進行する。従って、LLPS形成はウイルスによる宿主細胞のシャットオフ機構に大きく関連していると考えられるが、それだけに留まらず、LLPSは細胞の転写全体に関連している。従って、本研究によって、IE1依存的なLLPS形成機構の一端が解明されたことは、バキュロウイルスのシャットオフ機構研究が、細胞の転写機構全体を理解するための重要なモデルシステムと成り得ることを示している。

Research Products

• [Int'l Joint Research] Universiti Sains Malaysia(マレーシア)
• [Int'l Joint Research] Universiti Sains Malaysia(マレーシア)
• [Journal Article] Apoptotic arms races in insect‐baculovirus coevolution (2021)
  • Author(s): Nagamine Toshihiro
  • Journal Title: Physiological Entomology Volume: 47 Issue: 1 Pages: 1-10
  • DOI: 10.1111/phen.12371
  • Peer Reviewed / Open Access
• [Journal Article] Process development for quantitation and vaccine efficacy assessment of recombinant hemagglutinin-neuraminidase (2020)
  • Author(s): Lye Ping Ying、Noor Suriani Mohd、Shohaimi Syamsiah Aini、Junoh Niny Fariza、Tan Soo Choon、Iwamoto Shinichi、Kotani Eiji、Norazmi Mohd Nor、Nagamine Toshihiro、Mori Hajime、Liew Mervyn W.O.
  • Journal Title: Process Biochemistry Volume: 90 Pages: 204-214
  • DOI: 10.1016/j.procbio.2019.11.018
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Salt‐induced phase separation of CHAPS and potential application for hydrophobic molecule extraction (2019)
  • Author(s): Lian Ai A、Tan Soo C、Nagamine Toshihiro、Liew Mervyn WO
  • Journal Title: Journal of Chemical Technology & Biotechnology Volume: 印刷中 Issue: 7 Pages: 2284-2291
  • DOI: 10.1002/jctb.6016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A nuclear envelop-associated baculovirus protein promotes intranuclear lipid accumulation during infection (2019)
  • Author(s): Nagamine Toshihiro、Inaba Takehiko、Sako Yasushi
  • Journal Title: Virology Volume: 532 Pages: 108-117
  • DOI: 10.1016/j.virol.2019.04.006
  • Peer Reviewed / Open Access
• [Presentation] バキュロウイルスのODV（包埋ウイルス）形成機構：PR脂質によるカプシド捕獲モデル (2020)
  • Author(s): 永峰俊弘・佐甲靖志
  • Organizer: 蚕糸・昆虫機能利用学術講演会
• [Presentation] バキュロウイルス核膜タンパク質Bm5 / Ac13（BION）の機能解析 (2019)
  • Author(s): 永峰俊弘・稲葉岳彦・佐甲靖志
  • Organizer: 日本蚕糸学会
• [Presentation] バキュロウイルス核膜タンパク質Bm5/Ac13の局在機構解析 (2018)
  • Author(s): 永峰俊弘 稲葉岳彦 佐甲靖志
  • Organizer: 日本蚕糸学会
• [Book] 月刊「昆虫と自然」 (2022)
  • Author(s): 永峰俊弘
  • Total Pages: 4
  • Publisher: ニューサイエンス社
• [Book] アグリバイオ (2019, vol.3, No.12, 1149-1153) (2019)
  • Author(s): 永峰俊弘
  • Total Pages: 5
  • Publisher: 北隆館