Analysis of molecular mechanisms for apoptosis in insects infected with baculoviruses

杆状病毒感染昆虫细胞凋亡的分子机制分析

• 批准号: 14206007
• 负责人: KOBAYASHI Michihiro
• 金额: $ 34.28万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14206007/
• 关键词: 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

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

LD652Y细胞被七种不同的核型多发病毒（NPV）感染，包括来自加州Apotographa（ACMNPV），Bombyx Mori（BMNPV），Cunea（HycunPV），Spodoptera Exigua（Semnpv），lymania dispar和lymanta parta and lymantraia dispar（spodoptera spodoptera litura（spltmnpv）。结果表明，在凋亡中，被BMNPV，HycunPV，SemnPV，OPMNPV和SpltMNPV感染的LD652Y细胞。在感染了HycunPV的LD652Y细胞中，尽管没有可检测到的Budded病毒和多面蛋白的可检测到的产量，但仍考虑了一定量的病毒DNA。 Northern印迹和免疫印迹分析表明，凋亡3（HYCU-IAP3）的HycunPV抑制剂已显示在SF9细胞中起作用，在HycunPV感染的LD652Y细胞中表达了与HyCunPV感染的抗蛋白细胞相当的水平，而没有任何响应，而没有任何响应量，而没有任何响应。 These results imply that the relative ease of apoptosis in NPV-infected Ld652Y cells are l … More argely dependent on inherit cellular properties rather than functions of the respective NPVs, and indicate that the defect in progression in progression production is not merely due to the virus-induced apoptosis in HycuNPV-infected Ld652Y Cells.HycuNPV infection protected SpIm cells from放线菌素D诱导的凋亡早在感染后4小时。通过南部杂交分析表明，HycunPV基因组具有三个IAP基因成员，这些基因被指定为HYCU-IAP1，HYCU-IAP2和HYCU-IAP3，因为它们的氨基酸序列与其他细菌病毒中鉴定的IAPS具有氨基酸序列同源性。 SF9细胞中瞬态表达测定的HYCU-IAP的功能分析表明，放线肌蛋白D诱导的HYCU-IAP3阻塞凋亡并反应了p35缺乏症状氧化剂ACMNPV的复制，而HYCU-IAP1和HYCU-IAP2和HYCU-IAP2并未显示任何抗抑制性函数。在HycunPV感染允许的SPIM细胞感染期间，RNAi敲低HYCU-IAP3表达。这些结果表明，HYCU-IAP3对于允许性尖刺细胞的HycunPV感染期间的凋亡是必不可少的。较少的

项目成果

Abortive replication of Bombyx mori nucleopolyhyedrovirus in Sf9 and High Five cells : Defective nuclear transport of the virions

家蚕核多氢病毒在 Sf9 和 High Five 细胞中的复制失败：病毒颗粒的核运输缺陷

• 发表时间: 2006
• 期刊: Virology 347・2
• 作者: Katou;Y.;Ikeda;M.;Kobayashi;M.
• 通讯作者: M.

A recombinant Bombyx mori nucleopolyhyedrovirus possessing hrf-1 gene replicates in nonpermissive Lymantria dispar IPLB-652Y cell line

具有hrf-1基因的重组家蚕核多氢病毒在不允许的舞毒蛾IPLB-652Y细胞系中复制

• 发表时间: 2006
• 期刊: Journal of Insect Biotechnology and Sericology 75・1
• 作者: Ishikawa;H.;Ogasawara;T.;Ikeda;M.;Kobayashi;M.
• 通讯作者: M.

Accumulation of non-viral 45.5K and 44K polypeptides inversely correlates with that of viral structural polypeptides in the midgut of the silkworm, Bombyx mori

家蚕中肠中非病毒 45.5K 和 44K 多肽的积累与病毒结构多肽的积累呈负相关

• 发表时间: 2005
• 期刊: Journal of Insect Biotechnology and Sericology 74・2
• 作者: Sotoshiro;H.;Ikeda;M.;Tanaka;T.;Kobayashi;M.
• 通讯作者: M.

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Ishikawa, H., Ikeda, M., Yanagimoto, K.et al.: "Induction of apoptosis in IPLB-Ld 652Y cell line infected with nucleopolyhedroviruses"J.Geu.Virol.. 84・3. 705-714 (2003)

Ishikawa, H.、Ikeda, M.、Yanagimoto, K.等：“感染核多角体病毒的 IPLB-Ld 652Y 细胞系中的细胞凋亡的诱导”J.Geu.Virol.. 84・3 (2003)。