| Project/Area Number |
14206007
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
蚕糸・昆虫利用学
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| Research Institution | Nagoya University |
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Principal Investigator |
KOBAYASHI Michihiro Nagoya University, Graduate School of Bioagricultural Sciences, Professor, 大学院・生命農学研究科, 教授 (60111837)
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| Co-Investigator(Kenkyū-buntansha) |
IKEDA Motoko Nagoya University, Graduate School of Bioagricultural Sciences, Associate Professor, 大学院・生命農学研究科, 助教授 (20262892)
NIIMI Teruyuki Nagoya University, Graduate School of Bioagricultural Sciences, Assistant Professor, 大学院・生命農学研究科, 助手 (00293712)
河村 敏 岐阜県生物技術研究所, 専門研究員
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| Project Period (FY) |
2002 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥53,560,000 (Direct Cost: ¥41,200,000、Indirect Cost: ¥12,360,000)
Fiscal Year 2005: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2004: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2003: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2002: ¥27,820,000 (Direct Cost: ¥21,400,000、Indirect Cost: ¥6,420,000)
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| Keywords | nucleopolyhedrovirus / Ld652Y cell / fall webworm / gypsy moth / apoptosis / global protein synthesis shutdown / anti-viral responce / バキュロウイルス / 組換えウイルス / gp64 / 培養細胞 / sf9細胞 / High Five細胞 / 昆虫 |
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| Research Abstract |
Ld652Y cells were infected with seven different nucleopolyhedroviruses (NPVs), including those from Autographa californica (AcMNPV), Bombyx mori (BmNPV), Hyphantria cunea (HycuNPV), Spodoptera exigua (SeMNPV), Lymantria dispar, Orgyia pseudotsugata (OpMNPV) and Spodoptera litura (SpltMNPV). The results showed that Ld652Y cells infected with BmNPV, HycuNPV, SeMNPV, OpMNPV and SpltMNPV underwent apoptosis. In HycuNPV-infected Ld652Y cells, a considerable amount of viral DNA was synthesized although there was no detectable yield of budded virions and polyhedrin. Northern blot and immunoblot analyses revealed that HycuNPV inhibitor of apoptosis 3 (Hycu-IAP3), which has been shown to function in Sf9 cells, was expressed in HycuNPV-infected Ld652Y cells at a level comparable with that in HycuNPV-infected SpIm cells, which produced a high titer of progeny virions without any apoptotic response. These results imply that the relative ease of apoptosis induction in NPV-infected Ld652Y cells is l
… More
argely dependent on inherent cellular properties rather than functions of the respective NPVs, and indicate that the defect in progeny virion production is not merely due to the virus-induced apoptosis in HycuNPV-infected Ld652Y cells.
HycuNPV infection protected SpIm cells from actinomycin D-induced apoptosis as early as 4 h postinfection. Analysis by Southern hybridization revealed that the HycuNPV genome possessed three members of iap genes that were designated as hycu-iap1,hycu-iap2 and hycu-iap3 because of their amino acid sequence homology with iaps identified in other baculoviruses. Functional analysis of Hycu-IAPs by transient expression assay in Sf9 cells revealed that Hycu-IAP3 blocked apoptosis induced by actinomycin D and rescued replication of p35-deficient-mutant AcMNPV, while Hycu-IAP1 and Hycu-IAP2 did not show any anti-apoptotic functions. Knockdown of hycu-iap3 expression by RNAi during HycuNPV infection of permissive SpIm cells induced apoptosis. These results indicate that Hycu-IAP3 is essential for blockage of apoptosis during HycuNPV infection of permissive SpIm cells. Less
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