Visualization of the N-pro protein in living cells using biarsenically labeling tetracysteine-tagged classical swine fever virus

文献类型： 外文期刊

第一作者： Li, Yongfeng

作者： Li, Yongfeng;Shen, Liang;Li, Chao;Huang, Junhua;Zhao, Bibo;Sun, Yuan;Li, Su;Luo, Yuzi;Qiu, Hua-Ji

作者机构：

关键词： Classical swine fever virus;N-pro protein;Tetracysteine tag;Real-time fluorescence imaging

期刊名称：VIRUS RESEARCH （ 影响因子：3.303； 五年影响因子：3.445 ）

ISSN： 0168-1702

年卷期： 2014 年 189 卷

页码：

收录情况： SCI

摘要： Real-time fluorescence imaging of viral proteins in living cells is a valuable means to study virus-host interactions, and tetracysteine (TC)-biarsenical technology has been used in several viruses but not in classical swine fever virus (CSFV). Here, we generated CSFV mutants vSMTC385 or vSMTC412 bearing the small IC tag (CCPGCC) in the N-terminal region of the N-pro protein. The mutants showed growth characteristics indistinguishable from that of the wild-type virus, and retained similar N-pro subcellular localization to that of the parent virus. Furthermore, labeling with membrane-permeable biarsenical dye resulted in the fluorescent N-pro protein in the context of virus infection. Finally, we showed that N-pro was localized in the cytoplasm of CSFV-infected cells at 27 h post-infection (hpi) and present in the nucleus at 48 hpi, and the nuclear import and export was clearly observed from 36.5 to 37 hpi. Interestingly, our results demonstrated that N-pro transported across the nuclear pores by passive diffusion, which might be prevented by exogenous interferon regulatory factor 3 interacting with N-pro. Taken together, biarsenical labeling allows real-time visualization of the nucleus import and export of the fluorescent N-pro protein in CSFV-infected living cells. (C) 2014 Elsevier B.V. All rights reserved.

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