| Project/Area Number |
17076017
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Science and Engineering
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| Research Institution | Hosei University |
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Principal Investigator |
HONDA Ayae Hosei University, 生命科学部, 教授 (80343747)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Tadashi 東京大学, 医科学研究所, 教授 (40134621)
SAKAGUCHI Takemasa 広島大学, 医歯薬大学院, 教授 (70196070)
YASUDA Jiro 学警察研究所, 法科学第一部, 室長 (10282518)
SHIMIZU Kazufumi 日本大学, 医学部, 教授 (50004677)
IWATA Akira 日本生物科学研究所, 研究部, 主任研究員 (70193745)
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| Project Period (FY) |
2005 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥95,700,000 (Direct Cost: ¥95,700,000)
Fiscal Year 2009: ¥16,700,000 (Direct Cost: ¥16,700,000)
Fiscal Year 2008: ¥19,700,000 (Direct Cost: ¥19,700,000)
Fiscal Year 2007: ¥20,300,000 (Direct Cost: ¥20,300,000)
Fiscal Year 2006: ¥19,400,000 (Direct Cost: ¥19,400,000)
Fiscal Year 2005: ¥19,600,000 (Direct Cost: ¥19,600,000)
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| Keywords | インフルエンザウイルス / 光ピンセット / 宿主タンパク質 / Ebp1 / シアル酸 / RNAポリメラーゼ / 細胞周期 / 宿主因子 / 単一ウイルス / プロモーター / GFP / ErbB3 / 遺伝子発現誘導 / 宿主蛋白質 / 仙台ウイルス / 蛋白質修飾 / RNAi / 感染応答 / 単一細胞 |
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| Research Abstract |
1. To understand whether influenza virus infection is cell cycle dependent or not, we developed the microchip chamber system combined with optical trapping of single virus. Using this system, the fluorescently labeled influenza virus was successfully trapped under the lens using 1064 nm laser. The trapped influenza virus was transported on the dividing cell and released on it, but influenza virus did not attached. The same unattached virus on the dividing cell was trapped again and transported to the resting cell and released on it, the virus was attached on the resting cell. This result encouraged us to identify the difference between the resting and dividing cells. At first, we tried to compare the ・ 2,3-sialic acid contents between both cells. The result indicated that the content of ・ 2,3-sialic acid is higher in the resting cells than in the dividing cells.
2. Host protein Ebp1, ErbB3 binding protein, interacted with PB1 subunit of influenza virus RNA dependent RNA polymerase, and inhibited the RNA synthesis in vitro but not endonuclease associated with RNA dependent RNA polymerase. Interestingly Ebp1 expression was induced by influenza virus infection. To understand the induction mechanism of Ebp1 expression by influenza virus infection, the different combination among the eight viral protein expression plasmids and viral RNA (vRNA) expression plasmid were transfected into the cell and assayed the Ebp1 expression level using RT-PCR. The result indicated that the expression of vRNP complex, vRNA, influenza virus RNA dependent RNA polymerase and NP, influenced the Ebp1 expression. 3. Influenza virus infection induced changing of the cellular membrane strength.
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