Identification of protein-protein interactions of the occlusion-derived virus-associated proteins of Helicoverpa armigera nucleopolyhedrovirus.
Identification of protein-protein interactions of the occlusion-derived virus-associated proteins of Helicoverpa armigera nucleopolyhedrovirus.
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DOI：
10.1099/vir.0.017103-0
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发表时间：
2010-03
期刊：
影响因子：
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通讯作者：
Ke Peng;Minzhi Wu;F. Dèng;Jingjiao Song;Chunsheng Dong;Huálín Wáng;Zhìhóng Hú
中科院分区：
文献类型：
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作者：
Ke Peng;Minzhi Wu;F. Dèng;Jingjiao Song;Chunsheng Dong;Huálín Wáng;Zhìhóng Hú
本研究的目的是确定蛋白质之间的相互作用的吸藏衍生病毒（ODV）的棉铃虫核型多角体病毒（HearNPV），第二组α杆状病毒科。为了实现这一目标，选择了39个潜在的ODV结构蛋白的基因进行克隆，并在Gal 4酵母双杂交（Y2 H）系统中表达。直接交叉Y2 H测定鉴定了22种相互作用，包括13种二元相互作用[HA 9-ODV-EC 43、ODV-E56- 38 K、ODV-E56-PIF 3、LEF 3-解旋酶、LEF 3-碱性核酸酶（AN）、GP 41 - 38 K、GP 41-HA 90、38 K-PIF 3、38 K-PIF 2、VP 80-HA 100、ODV-E66-PIF 3、ODV-E66-PIF 2和PIF 3-PIF 2]和9种自缔合（IE 1、HA 44、LEF 3、HA 66、GP 41、CG 30、38 K、PIF 3和P24）。这些相互作用中的五个-LEF 3-解旋酶和LEF 3-AN，以及IE 1，LEF 3和38 K的自缔合-先前已在苜蓿银纹夜蛾多核型多角体病毒中报道。由于HA 44和HA 100是两个新发现的II组ODV蛋白，进一步证实了它们之间的相互作用。HA 44的自结合通过His pull-down测定来验证，并且VP 80-HA 100的相互作用通过免疫共沉淀测定来证实。本文综述了杆状病毒与45种病毒蛋白和5种宿主蛋白的68种相互作用，为深入了解杆状病毒感染的分子机制提供了参考。
The purpose of this study was to identify protein-protein interactions among the components of the occlusion-derived virus (ODV) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV), a group II alphabaculovirus in the family Baculoviridae. To achieve this, 39 selected genes of potential ODV structural proteins were cloned and expressed in the Gal4 yeast two-hybrid (Y2H) system. The direct-cross Y2H assays identified 22 interactions comprising 13 binary interactions [HA9-ODV-EC43, ODV-E56-38K, ODV-E56-PIF3, LEF3-helicase, LEF3-alkaline nuclease (AN), GP41-38K, GP41-HA90, 38K-PIF3, 38K-PIF2, VP80-HA100, ODV-E66-PIF3, ODV-E66-PIF2 and PIF3-PIF2] and nine self-associations (IE1, HA44, LEF3, HA66, GP41, CG30, 38K, PIF3 and P24). Five of these interactions - LEF3-helicase and LEF3-AN, and the self-associations of IE1, LEF3 and 38K - have been reported previously in Autographa californica multiple nucleopolyhedrovirus. As HA44 and HA100 were two newly identified ODV proteins of group II viruses, their interactions were further confirmed. The self-association of HA44 was verified with a His pull-down assay and the interaction of VP80-HA100 was confirmed by a co-immunoprecipitation assay. A summary of the protein-protein interactions of baculoviruses reported so far, comprising 68 interactions with 45 viral proteins and five host proteins, is presented, which will facilitate our understanding of the molecular mechanisms of baculovirus infection.