Probing the nuclear import signal and nuclear transport molecular determinants of PRV ICP22.
Probing the nuclear import signal and nuclear transport molecular determinants of PRV ICP22.
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探讨 PRV ICP22 的核输入信号和核运输分子决定因素
DOI：
10.1186/s13578-016-0069-7
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发表时间：
2016
影响因子：
7.5
通讯作者：
Li M
中科院分区：
文献类型：
--
作者：
Cai M;Jiang S;Zeng Z;Li X;Mo C;Yang Y;Chen C;Xie P;Bian Y;Wang J;Huang J;Chen D;Peng T;Li M
单纯疱疹病毒1型（HSV - 1）ICP22是一种多功能蛋白，对HSV - 1的复制非常重要。伪狂犬病病毒（PRV）ICP22（P - ICP22）是HSV - 1 ICP22的同源物，据报道它能够选择性地改变PRV不同动力学类别基因的转录，然而，其亚细胞定位、定位信号以及为执行此功能而进行运输的分子决定因素却鲜为人知。
在本研究中，通过利用活细胞荧光显微镜技术，与增强型黄色荧光蛋白（EYFP）基因融合的P - ICP22在活细胞中瞬时表达，结果显示在没有其他病毒蛋白的情况下，其主要定位于细胞核。通过转染一系列与EYFP融合的P - ICP22缺失突变体，首次确定了P - ICP22真正的核定位信号（NLS）及其关键氨基酸（aa），并分别将其定位到氨基酸41 - 60（PASTPTPPKRGRYVVEHPEY）和氨基酸49 - 50（KR）。此外，还证明了P - ICP22是通过Ran、输入蛋白α1和α7介导的途径被靶向到细胞核的。
我们在此报告的研究结果揭示了P - ICP22核运输的NLS和分子机制，这些结果将为描绘P - ICP22在PRV感染过程中的生物学作用开辟新的途径。
BackgroundHerpes simplex virus 1 (HSV-1) ICP22 is a multifunctional protein and important for HSV-1 replication. Pseudorabies virus (PRV) ICP22 (P-ICP22) is a homologue of HSV-1 ICP22 and is reported to be able to selectively modify the transcription of different kinetic classes of PRV genes, however, the subcellular localization, localization signal and molecular determinants for its transport to execute this function is less well understood.ResultsIn this study, by utilizing live cells fluorescent microscopy, P-ICP22 fused to enhanced yellow fluorescent protein (EYFP) gene was transient expressed in live cells and shown to exhibit a predominantly nucleus localization in the absence of other viral proteins. By transfection of a series of P-ICP22 deletion mutants fused to EYFP, a bona fide nuclear localization signal (NLS) and its key amino acids (aa) of P-ICP22 was, for the first time, determined and mapped to aa 41–60 (PASTPTPPKRGRYVVEHPEY) and aa 49–50 (KR), respectively. Besides, the P-ICP22 was demonstrated to be targeted to the nucleus via Ran-, importin α1-, and α7-mediated pathway.ConclusionsOur findings reported herein disclose the NLS and molecular mechanism for nuclear transport of P-ICP22, these results will uncover new avenues for depicting the biological roles of P-ICP22 during PRV infection.