权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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

基本信息

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

项目摘要

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

用来自苜蓿银纹夜蛾（AcMNPV）、桑尺蠖（BmNPV）、美国白蛾（HycuNPV）、甜菜夜蛾（SeMNPV）、舞毒蛾（Lymantria dispar）、拟杉毒蛾（Orgyia pseudotsugata）和斜纹夜蛾（Spodoptera litura）的7种不同的核型多角体病毒（NPVs）感染Ld 652 Y细胞。结果表明，BmNPV、HycuNPV、SeMNPV、OpMNPV和SpltMNPV感染的Ld 652 Y细胞发生凋亡。在HycuNPV感染的Ld 652 Y细胞中，合成了相当数量的病毒DNA，尽管没有可检测到的出芽病毒体和多角体蛋白的产量。北方印迹和免疫印迹分析显示，HycuNPV凋亡抑制剂3（Hycu-IAP 3），这已被证明是在Sf 9细胞中的功能，在HycuNPV感染的Ld 652 Y细胞中的表达水平与HycuNPV感染的SpIm细胞中的表达水平相当，这产生了高滴度的子代病毒粒子没有任何凋亡反应。这些结果表明，在HPV感染的Ld 652 Y细胞中，诱导凋亡的相对容易性为10%。 ...更多信息 HycuNPV感染Ld 652 Y细胞后，其子代病毒体产生的缺陷不仅仅是由于HycuNPV诱导的细胞凋亡，HycuNPV感染SpIm细胞后4 h即可保护细胞免受放线菌素D诱导的细胞凋亡的影响。Southern杂交分析表明，HycuNPV基因组中含有3个iap基因，由于其氨基酸序列与其它杆状病毒的iap基因同源，分别命名为hycu-iap 1、hycu-iap 2和hycu-iap 3。在Sf 9细胞中瞬时表达Hycu-IAP 3，Hycu-IAP 1和Hycu-IAP 2不具有抗细胞凋亡的功能，Hycu-IAP 3能阻断放线菌素D诱导的细胞凋亡，并能挽救p35缺陷型AcMNPV的复制。在HycuNPV感染允许的SpIm细胞期间通过RNAi敲低hycu-iap 3表达诱导细胞凋亡。这些结果表明，Hycu-IAP 3对于HycuNPV感染许可SpIm细胞期间阻断细胞凋亡至关重要。少