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目的 鸭瘟病毒（Duck plague virus, DPV）属于α疱疹病毒亚科，其编码的UL6蛋白是疱疹病毒门蛋白的同源物，在细胞核中参与病毒衣壳的组装和基因组的包装，其在细胞中的正确定位对于功能的发挥至关重要。本研究旨在系统性研究DPV UL6蛋白在细胞中的亚细胞定位特征，分析其潜在的核定位信号（Nuclear localization signal,NLS）和关键氨基酸，解析DPV UL6蛋白的入核机制。方法 本文主要运用间接免疫荧光法通过检测与DPV UL6蛋白及其突变体融合表达的Flag标签来分析UL6蛋白和UL6突变体的亚细胞定位。结果 DPV UL6蛋白中存在两个经典的核定位信号基序：一个双分型核定位信号（Bipartite NLS, bNLS）和一个单分型核定位信号（Monopartite NLS, mNLS），并且二者在介导UL6蛋白入核的过程中具有协同作用。进一步的氨基酸突变分析发现，UL6 K142A、UL6 K737A、UL6 R738A和UL6 R740A突变体影响UL6蛋白进入细胞核。结论bNLS和mNLS对DPV UL6蛋白的核定位均至关重要，两者的协同作用是实现其核定位所必需的，且R738和R740为DPV UL6蛋白核定位的必需氨基酸。
Abstract：Objective Duck plague virus(DPV) is a member of the Alphaherpesvirinae subfamily. The UL6 protein encodes by DPV is a homolog of the herpesviral portal protein and participates in viral capsid assembly and genome packaging within the nucleus. Proper subcellular localization of UL6 is therefore essential for its biological function. This study aims to systematically investigate the subcellular localization of DPV UL6 protein, identify its potential nuclear localization signals and key amino acid residues, and elucidate the molecular mechanism underlying UL6 nuclear import. Methods Indirect immunofluorescence assays were employed to analyze the subcellular localization of the DPV UL6 protein and its mutant forms by detecting Flagtagged UL6 and corresponding mutants expressed in cells. Results Two classical nuclear localization signal motifs were identified within DPV UL6 protein: a bipartite nuclear localization signal(bNLS) and a monopartite nuclear localization signal(mNLS). These two motifs act synergistically to mediate the nuclear import of UL6. Further site-directed mutagenesis revealed that the UL6 K142A, UL6 K737A, UL6 R738A, and UL6 R740A mutants exhibited impaired nuclear localization.Conclusion Both the bNLS and mNLS are essential for the nuclear localization of the DPV UL6 protein, and their synergistic action is required for efficient nuclear import. In particular, residues R738 and R740 are indispensable for UL6 nuclear targeting.
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基本信息:
中图分类号:S852.65
引用信息:
[1]杨静,宋迪,刘思佳,等.双重核定位信号协同作用促进鸭瘟病毒UL6蛋白入核[J].病毒学报().
基金信息:
四川省科技计划项目（项目号：2020YJ0395）,题目：鸭瘟病毒pUL6的多核定位信号对其入核的调控以及对病毒复制的影响
2026-02-26
2026-02-26
2026-02-26