A study of the Autographa californica multiple nucleopolyhedrovirus ODV envelope protein p74 using a GFP tag.
A study of the Autographa californica multiple nucleopolyhedrovirus ODV envelope protein p74 using a GFP tag.
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DOI：
10.1099/0022-1317-82-9-2279
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发表时间：
2001-09
期刊：
影响因子：
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通讯作者：
J. Slack;Edward M. Dougherty;S. Lawrence
中科院分区：
文献类型：
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作者：
J. Slack;Edward M. Dougherty;S. Lawrence
加州签名虫多核多角体病毒（AcMNPV）蛋白p74与闭塞性衍生病毒（ODV）包膜相关。p74对于ODV的口腔感染至关重要，并被认为在中肠附着和/或融合中发挥作用。本研究将p74蛋白与绿色荧光蛋白（GFP）在框架内表达，构建p74-GFP嵌合体。嵌合体的c端GFP部分有助于可视化p74在杆状病毒感染的frugiperda Spodoptera （Sf-9）细胞中的运输。p74-GFP嵌合蛋白定位于细胞核的核内环区，并被发现与细胞裂解物的微泡部分共沉淀。一系列截断的p74在重组杆状病毒中表达为p74- gfp嵌合体。当从p74中删除c端区域S580-F645后，p74- gfp嵌合体定位变得非特异性，嵌合体变得可溶。p74区域S580-F645以与全长p74相似的模式将GFP定向到核内环区。p74的疏水C端在蛋白质定位中起作用，并可能在跨膜锚定和插入中起作用。
The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) protein p74 is associated with the occlusion-derived virus (ODV) envelope. p74 is essential for oral infectivity of ODV and has been proposed to play a role in midgut attachment and/or fusion. In this study, p74 protein was expressed in-frame with green fluorescent protein (GFP) to create a p74-GFP chimera. The C-terminal GFP portion of the chimera facilitated visualization of the trafficking of p74 in baculovirus-infected Spodoptera frugiperda (Sf-9) cells. p74-GFP chimeric proteins localized in the intranuclear ring zone of the nucleus and were found to co-precipitate with the microvesicle fraction of cell lysates. A series of truncations of p74 was expressed as p74-GFP chimeras in recombinant baculoviruses. When C-terminal region S580-F645 was deleted from p74, p74-GFP chimera localization became non-specific and chimeras became soluble. p74 region S580-F645 directed GFP to the intranuclear ring zone in a similar pattern to full-length p74. The hydrophobic C terminus of p74 plays a role in protein localization and possibly in transmembrane anchoring and insertion.