Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins
Intracellular localization of Crimean-Congo Hemorrhagic Fever (CCHF) virus glycoproteins
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DOI：
10.1186/1743-422x-2-42
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发表时间：
2005-01-01
期刊：
影响因子：
4.8
通讯作者：
Flick, Ramon
中科院分区：
文献类型：
--
作者：
Haferkamp, Sebastian;Fernando, Lisa;Flick, Ramon
背景资料：克里米亚-刚果出血热病毒（CCHFV）是布尼亚病毒科Nairovirus属成员，是一种蜱传病原体，可引起人类严重疾病。为了更好地了解CCHFV的生命周期并探索潜在的干预策略，我们研究了由M基因组片段编码的糖蛋白的生物合成和细胞内靶向。在测定CCHFV参考株IbAr 10200的全基因组序列后，我们产生了用于单独表达糖蛋白G（N）和G（C）的表达质粒，使用CMV和鸡β-肌动蛋白驱动的启动子。重组表达的CCHFV糖蛋白的细胞定位进行了比较，在病毒感染过程中，使用间接免疫荧光测定，亚细胞分级分离/蛋白质印迹分析和共聚焦显微镜真实的糖蛋白。为了进一步阐明两种糖蛋白的潜在细胞内靶向/保留信号，分析了含有CCHFV糖蛋白不同部分的GFP融合蛋白的细胞内靶向。N-末端糖蛋白G（N）定位于高尔基复合体，这是由该蛋白质的胞质结构域和胞外域中的保留/靶向信号介导的过程。相反，C-末端糖蛋白G（C）保留在内质网中，但可以通过G（N）的共表达被拯救到高尔基复合体中。结论：这些数据与大多数布尼亚病毒糖蛋白的胞内靶向一致，支持布尼亚病毒颗粒在高尔基体中组装和出芽的一般模型。
Background: Crimean-Congo Hemorrhagic Fever virus (CCHFV), a member of the genus Nairovirus, family Bunyaviridae, is a tick-borne pathogen causing severe disease in humans. To better understand the CCHFV life cycle and explore potential intervention strategies, we studied the biosynthesis and intracellular targeting of the glycoproteins, which are encoded by the M genome segment.Results: Following determination of the complete genome sequence of the CCHFV reference strain IbAr10200, we generated expression plasmids for the individual expression of the glycoproteins G(N) and G(C), using CMV- and chicken beta-actin-driven promoters. The cellular localization of recombinantly expressed CCHFV glycoproteins was compared to authentic glycoproteins expressed during virus infection using indirect immunofluorescence assays, subcellular fractionation/western blot assays and confocal microscopy. To further elucidate potential intracellular targeting/retention signals of the two glycoproteins, GFP-fusion proteins containing different parts of the CCHFV glycoprotein were analyzed for their intracellular targeting. The N-terminal glycoprotein G(N) localized to the Golgi complex, a process mediated by retention/targeting signal(s) in the cytoplasmic domain and ectodomain of this protein. In contrast, the C-terminal glycoprotein G(C) remained in the endoplasmic reticulum but could be rescued into the Golgi complex by co-expression of G(N).Conclusion: The data are consistent with the intracellular targeting of most bunyavirus glycoproteins and support the general model for assembly and budding of bunyavirus particles in the Golgi compartment.