Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
J Virol. 2013 Aug;87(15):8342-50. doi: 10.1128/JVI.03549-12. Epub 2013 May 22.
For most parainfluenza viruses, a virus type-specific interaction between the hemagglutinin-neuraminidase (HN) and fusion (F) proteins is a prerequisite for mediating virus-cell fusion and cell-cell fusion. The molecular basis of this functional interaction is still obscure partly because it is unknown which region of the F protein is responsible for the physical interaction with the HN protein. Our previous cell-cell fusion assay using the chimeric F proteins of parainfluenza virus 5 (PIV5) and simian virus 41 (SV41) indicated that replacement of two domains in the head region of the PIV5 F protein with the SV41 F counterparts bestowed on the PIV5 F protein the ability to induce cell-cell fusion on coexpression with the SV41 HN protein while retaining its ability to induce fusion with the PIV5 HN protein. In the study presented here, we furthered the chimeric analysis of the F proteins of PIV5 and SV41, finding that the PIV5 F protein could be converted to an SV41 HN-specific chimeric F protein by replacing five domains in the head region with the SV41 F counterparts. The five SV41 F-protein-derived domains of this chimera were then divided into 16 segments; 9 out of 16 proved to be not involved in determining its specificity for the SV41 HN protein. Finally, mutational analyses of a chimeric F protein, which harbored seven SV41 F-protein-derived segments, revealed that replacement of at most 21 amino acids of the PIV5 F protein with the SV41 F-protein counterparts was enough to convert its HN protein specificity.
对于大多数副黏病毒而言,血凝素-神经氨酸酶(HN)和融合(F)蛋白之间的病毒类型特异性相互作用是介导病毒-细胞融合和细胞-细胞融合的前提。这种功能相互作用的分子基础仍然不清楚,部分原因是未知 F 蛋白的哪个区域负责与 HN 蛋白发生物理相互作用。我们之前使用副流感病毒 5(PIV5)和猿猴病毒 41(SV41)的嵌合 F 蛋白进行的细胞-细胞融合测定表明,用 SV41 F 蛋白的两个结构域替换 PIV5 F 蛋白头部区域的两个结构域,可使 PIV5 F 蛋白在与 SV41 HN 蛋白共表达时获得诱导细胞-细胞融合的能力,同时保留其与 PIV5 HN 蛋白诱导融合的能力。在本研究中,我们进一步对 PIV5 和 SV41 的 F 蛋白进行嵌合分析,发现 PIV5 F 蛋白可以通过用 SV41 F 蛋白的五个结构域替换头部区域的五个结构域,转化为 SV41 HN 特异性嵌合 F 蛋白。该嵌合体的五个源自 SV41 F 蛋白的结构域然后被分成 16 个片段;其中 9 个片段被证明不参与决定其对 SV41 HN 蛋白的特异性。最后,对一个嵌合 F 蛋白进行突变分析,该嵌合 F 蛋白含有七个源自 SV41 F 蛋白的片段,结果表明,用 SV41 F 蛋白的最多 21 个氨基酸替换 PIV5 F 蛋白足以改变其 HN 蛋白的特异性。