Functional analysis of lipid domains in influenza virus infection with super-resolution microscopy

• Project/Area Number: 15K07957
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: ABE MITSUHIRO 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 (90415068)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: インフルエンザウイルス / フリップフロップ / 脂質ラフト / スフィンゴミエリン / PIP2 / 1分子イメージング / STORM / 脂質の非対称性 / インフルエンザ

Outline of Final Research Achievements

Analysis with a combination of lipid probes and super-resolution microscopy indicates that the influenza virus buds from the edges of the sphingomyelin(SM)/cholesterol(Chol) domains in MDCK cells. Several lines of evidence suggest that breakage of lipid asymmetry at the plasma membrane leads to the decrease of influenza virus proliferation. I screened for the factor involved in SM/Chol asymmetry at the plasma membrane. Immuno-electron microscopy with sodium dodecyl sulfate (SDS)-treated freeze-fracture replica labeling (SDS-FRL) indicates that the domains of phosphatidylinositol 4,5-bisphosphate (PIP2) increase in the inner leaflet of the plasma membrane. These results suggest that the stability of domains of PIP2 increases the influenza virus proliferation in mammalian cells.

Academic Significance and Societal Importance of the Research Achievements

本研究により、宿主側の細胞膜における脂質の非対称性は、インフルエンザウイルスの増殖・拡散に関わっていることが示唆された。同様の現象が、ほかのウイルスでも見られることが十分期待される。ウイルスは変異しやすく、構成タンパク質の分類上における種類の多さから、ウイルスそのものを標的とする薬剤の開発は難しい。本申請課題での結果を基に、ホスト側の因子を標的とした薬剤で、脂質の非対称性を不安定にし、ウイルスの感染効率が低下するものがあれば、ウイルスの増殖・拡散に対する薬剤の開発を容易に行えることが期待される。

Research Products

• [Int'l Joint Research] CNRS/Faculte de Pharmacie/Universite de Strasbourg(フランス)
• [Journal Article] Single-molecule diffusion-based estimation of ligand effects on G protein?coupled receptors (2018)
  • Author(s): Yanagawa Masataka、Hiroshima Michio、Togashi Yuichi、Abe Mitsuhiro、Yamashita Takahiro、Shichida Yoshinori、Murata Masayuki、Ueda Masahiro、Sako Yasushi
  • Journal Title: Science Signaling Volume: 11 Issue: 548 Pages: 1-16
  • DOI: 10.1126/scisignal.aao1917
  • Peer Reviewed / Open Access
• [Journal Article] Mutation-Specific Mechanisms of Hyperactivation of Noonan Syndrome SOS Molecules Detected with Single-molecule Imaging in Living Cells (2017)
  • Author(s): Nakamura Yuki、Umeki Nobuhisa、Abe Mitsuhiro、Sako Yasushi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 1-10
  • DOI: 10.1038/s41598-017-14190-6
  • Peer Reviewed / Open Access
• [Journal Article] Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs (2017)
  • Author(s): Masanao Kinoshita, Kenichi GN Suzuki, Nobuaki Matsumori, Misa Takada, Hikaru Ano, Kenichi Morigaki, Mitsuhiro Abe, Asami Makino, Toshihide Kobayashi, Koichiro M Hirosawa, Takahiro K Fujiwara, Akihiro Kusumi, Michio Murata
  • Journal Title: J Cell Biol Volume: 未定 Issue: 4 Pages: 1183-1204
  • DOI: 10.1083/jcb.201607086
  • NAID: 120005998045
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Dynamics of sphingomyelin- and cholesterol-enriched lipid domains during cytokinesis (2017)
  • Author(s): Abe M, Kobayashi T.
  • Journal Title: Methods in Cell Biology Volume: 137 Pages: 15-24
  • DOI: 10.1016/bs.mcb.2016.03.030
  • ISBN: 9780128096734
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Psychosine-triggered endomitosis is modulated by membrane sphingolipids through regulation of phosphoinositide 4,5-bisphosphate production at the cleavage furrow. (2016)
  • Author(s): Watanabe H, Okahara K, Naito-Matsui Y, Abe M, Go S, Inokuchi J, Okazaki T,Kobayashi T, Kozutsumi Y, Oka S, Takematsu H.
  • Journal Title: Mol Biol Cell. Volume: 27 Issue: 13 Pages: 2037-2050
  • DOI: 10.1091/mbc.e15-08-0555
  • Peer Reviewed / Open Access
• [Journal Article] Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation. (2016)
  • Author(s): Inaba T, Kishimoto T, Murate M, Tajima T, Sakai S, Abe M, Makino A, Tomishige N, Ishitsuka R, Ikeda Y, Takeoka S, Kobayashi T.
  • Journal Title: Proc Natl Acad Sci USA Volume: 113 Issue: 28 Pages: 7834-7839
  • DOI: 10.1073/pnas.1603513113
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A novel spingomyeling/cholesterol specific probe reveals the dynamics of the membrane domain during virus and in Nieman-Pick type C (2016)
  • Author(s): Makino, A., Sako, Y. et al.
  • Journal Title: FASEB J. Volume: 31 Issue: 4 Pages: 1301-1322
  • DOI: 10.1096/fj.201500075r
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Acute accumulation of free cholesterol induces the degradation of perilipin 2 and Rab18-dependent fusion of ER and lipid droplets in cultured human hepatocytes. (2016)
  • Author(s): Makino A, Hullin-Matsuda F, Murate M, Abe M, Tomishige N, Fukuda M, Yamashita S, Fujimoto T, Vidal H, Lagarde M, Delton I, Kobayashi T.
  • Journal Title: Mol Biol Cell Volume: 27 Issue: 21 Pages: 3293-3304
  • DOI: 10.1091/mbc.e15-10-0730
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Zds1/Zds2-PP2ACdc55 complex specifies signaling output from Rho1 GTPase (2016)
  • Author(s): Jonasson EM, Rossio V, Hatakeyama R, Abe M, Ohya Y, Yoshida S
  • Journal Title: J Cell Biol Volume: 212(1) Issue: 1 Pages: 51-61
  • DOI: 10.1083/jcb.201508119
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates (2015)
  • Author(s): Bhat HB, Ishitsuka R, Inaba T, Murate M, Abe M, Makino A, Kohyama-Koganeya A, Nagao K, Kurahashi A, Kishimoto T, Tahara M, Yamano A, Nagamune K, Hirabayashi Y, Juni N, Umeda M, Fujimori F, Nishibori K, Yamaji-Hasegawa A, Greimel P, Kobayashi T.
  • Journal Title: FASEB J Volume: 29(9) Issue: 9 Pages: 3920-34
  • DOI: 10.1096/fj.15-272112
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transbilayer distribution of lipids at nano scale. (2015)
  • Author(s): Murate M, Abe M, Kasahara K, Iwabuchi K, Umeda M, Kobayashi T
  • Journal Title: J Cell Sci Volume: 128 Pages: 1627-1638
  • DOI: 10.1242/jcs.163105
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 脂質イメージングによる脂質ラフトの解析 (2016)
  • Author(s): 阿部充宏
  • Organizer: 第42回レーザ顕微鏡研究会講演会
  • Place of Presentation: 理化学研究所 和光事業所 （埼玉県和光市）
  • Year and Date: 2016-07-07
• [Remarks] 細胞膜の受容体1分子の動きから薬効を評価
  • URL: http://www.riken.jp/pr/press/2018/20180919_1/
• [Remarks] マイタケ由来タンパク質がインフルエンザウイルスの増殖を抑制
  • URL: http://www.riken.jp/pr/press/2016/20160822_2/