Mutations in the DI-DII Linker of Human Parainfluenza Virus Type 3 Fusion Protein Result in Diminished Fusion Activity
Mutations in the DI-DII Linker of Human Parainfluenza Virus Type 3 Fusion Protein Result in Diminished Fusion Activity
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人副流感病毒 3 型融合蛋白 DI-DII 连接子突变导致融合活性降低
DOI：
10.1371/journal.pone.0136474
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发表时间：
2015-08
期刊：
影响因子：
3.7
通讯作者：
Zhiyu Wang
中科院分区：
文献类型：
--
作者：
Zhao L;Song Y;Sun C;Zhiyu Wang
人副流感病毒3型（HPIV 3）可导致婴幼儿严重呼吸道疾病，但目前尚无获得许可的疫苗或抗病毒剂可用于治疗。病毒和靶细胞膜的融合是其进入宿主细胞的先决条件，并由融合（F）蛋白直接介导。虽然已知F的几个结构域对调节膜融合活性具有重要作用，但HPIV 3 F蛋白的DI-DII接头（残基369-374）在融合性中的作用仍然不明确。为了便于我们理解该结构域可能在F诱导的细胞-细胞融合中发挥的作用，通过定点诱变将9个单突变工程化到该结构域中。采用牛痘病毒-T7 RNA聚合酶瞬时表达系统表达野生型或突变型F蛋白。这些突变体进行了分析，膜融合活性，细胞表面表达，F和HN蛋白之间的相互作用。该结构域中的每个突变的F蛋白具有与野生型F相似的细胞表面表达水平。所有这些都导致膜融合的所有步骤中的融合活性的显著降低。此外，所有这些融合缺陷突变体减少了细胞表面的HN-F复合物的量。总之，我们的工作结果表明，该区域对F.
Human parainfluenza virus type 3 (HPIV3) can cause severe respiratory tract diseases in infants and young children, but no licensed vaccines or antiviral agents are currently available for treatment. Fusing the viral and target cell membranes is a prerequisite for its entry into host cells and is directly mediated by the fusion (F) protein. Although several domains of F are known to have important effects on regulating the membrane fusion activity, the roles of the DI-DII linker (residues 369–374) of the HPIV3 F protein in the fusogenicity still remains ill-defined. To facilitate our understanding of the role of this domain might play in F-induced cell-cell fusion, nine single mutations were engineered into this domain by site-directed mutagenesis. A vaccinia virus-T7 RNA polymerase transient expression system was employed to express the wild-type or mutated F proteins. These mutants were analyzed for membrane fusion activity, cell surface expression, and interaction between F and HN protein. Each of the mutated F proteins in this domain has a cell surface expression level similar to that of wild-type F. All of them resulted in a significant reduction in fusogenic activity in all steps of membrane fusion. Furthermore, all these fusion-deficient mutants reduced the amount of the HN-F complexes at the cell surface. Together, the results of our work suggest that this region has an important effect on the fusogenic activity of F.
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影响因子：
64.8
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