Identification of the Nuclear Localization Signal Region of Duck Enteritis Virus UL14 and Its Interaction with VP16
Identification of the Nuclear Localization Signal Region of Duck Enteritis Virus UL14 and Its Interaction with VP16
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DOI：
10.1159/000452711
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发表时间：
2017-02
期刊：
影响因子：
4.6
通讯作者：
Fangjie Li;Yuwei Zhang;Shun Chen;Mingshu Wang;R. Jia;Dekang Zhu;Mafeng Liu;K. Sun;Qiao Yang;Ying Wu;Xiaoyue Chen;A. Cheng
中科院分区：
文献类型：
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作者：
Fangjie Li;Yuwei Zhang;Shun Chen;Mingshu Wang;R. Jia;Dekang Zhu;Mafeng Liu;K. Sun;Qiao Yang;Ying Wu;Xiaoyue Chen;A. Cheng
目的：鸭肠炎病毒（DEV）是α疱疹病毒亚科的一种病毒。VP 16和pUL 14均为DEV的被膜蛋白。研究方法：采用间接免疫荧光法（IFA）初步研究了病毒复制过程中pUL 14和VP 16在鸭胚成纤维细胞（DEF）中的共定位。在转染的DEFs中检测到pUL 14和VP 16的共表达。使用双分子荧光互补（BiFC）测定来确认pUL 14和VP 16之间的直接相互作用。为了更好地表征pUL 14的核定位结构域，我们设计了一系列缺失突变体，并将它们用VP 16转染。结果如下：我们的IFA结果表明，pUL 14在细胞质中与VP 16结合，并且pUL 14在DEV复制期间引导VP 16输入到细胞核中。BiFC测定揭示了pUL 14和VP 16复合物的存在。此外，pUL 14 N端1-98氨基酸（aa）在核定位信号（NLS）区发挥作用，促进VP 16易位入核，完成病毒的生活史。结论：我们的研究结果表明pUL 14可以将VP 16转运到细胞核中，并且N端1-98 aa可能含有pUL 14的NLS结构域。
Object: Duck enteritis virus (DEV) is a member of the Alphaherpesvirinae viruses. VP16 and pUL14 are both predicted to be tegument proteins of DEV. Methods: An indirect immunofluorescence assay (IFA) was performed for preliminary analysis of the colocalization of pUL14 and VP16, which detected their subcellular localization in duck embryo fibroblasts (DEFs) during virus replication. The coexpression of pUL14 and VP16 was detected in transfected DEFs. A bimolecular fluorescence complementation (BiFC) assay was used to confirm a direct interaction between pUL14 and VP16. To better characterize the nuclear localization domain of pUL14, we designed a series of deletion mutants and transfected them with VP16. Results: Our IFA findings indicated that pUL14 binds to VP16 in the cytoplasm and that pUL14 leads VP16 import into the nucleus during DEV replication. The BiFC assay revealed the presence of pUL14 and VP16 complexes. Furthermore, 1-98 amino acid (aa) at the N-terminus of pUL14 played a role in the nuclear localization signal (NLS) region and promoted translocation of VP16 into the nucleus to complete the virus life cycle. Conclusions: Our findings indicated that pUL14 could transport VP16 into the nucleus and that the N-terminal 1-98 aa may contain the NLS domain of pUL14.