Role of the C-terminal region of vervet monkey polyomavirus 1 VP1 in virion formation.
Role of the C-terminal region of vervet monkey polyomavirus 1 VP1 in virion formation.
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DOI：
10.1292/jvms.13-0568
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发表时间：
2014-05
期刊：
影响因子：
--
通讯作者：
Orba Y
中科院分区：
文献类型：
--
作者：
Yamaguchi H;Kobayashi S;Maruyama J;Sasaki M;Takada A;Kimura T;Sawa H;Orba Y
最近，我们在一只红毛猴体内检测到了新的红毛猴多瘤病毒1(VmPyV)。在其他多瘤病毒的主要衣壳蛋白VP1s的氨基酸序列中，VmPyV VP1是最长的，在C末端有额外的氨基酸残基。为了研究VmPyV VP1在病毒粒子形成中的作用，我们产生了VmPyV VP1的病毒样颗粒(VLP)，因为VLP是研究多瘤病毒病毒粒子形态特征的有用工具。将VmPyV VP1全长亚克隆到哺乳动物表达载体中，再将其导入人胚胎肾293T细胞。然后，用蔗糖梯度沉淀法从转基因细胞裂解产物中纯化VmPyV VLP。电子显微镜分析表明，VmPyV VP1形成直径约50 nm的VLP，仅定位于细胞核。此外，我们还构建了缺失C端116个氨基酸残基的缺失突变体VmPyV VP1(ΔC VP1)，并与野生型VmPyV VP1(WT VP1)的VLP形成效率和形态进行了比较。在表达VP1HEK293T细胞中，WT和ΔC VP1VLP的大小相似，但ΔCVP1VLP的数量明显低于WTVP1VLP。这些结果表明，VP1的长度与病毒粒子的形态无关，但VmPyV VP1的C-末端区域影响其VLP的形成效率。
Recently, we detected novel vervet monkey polyomavirus 1 (VmPyV) in a vervet monkey. Among amino acid sequences of major capsid protein VP1s of other polyomaviruses, VmPyV VP1 is the longest with additional amino acid residues in the C-terminal region. To examine the role of VmPyV VP1 in virion formation, we generated virus-like particles (VLPs) of VmPyV VP1, because VLP is a useful tool for the investigation of the morphological characters of polyomavirus virions. After the full-length VmPyV VP1 was subcloned into a mammalian expression plasmid, the plasmid was transfected into human embryonic kidney 293T (HEK293T) cells. Thereafter, VmPyV VLPs were purified from the cell lysates of the transfected cells via sucrose gradient sedimentation. Electron microscopic analyses revealed that VmPyV VP1 forms VLPs with a diameter of approximately 50 nm that are exclusively localized in cell nuclei. Furthermore, we generated VLPs consisting of the deletion mutant VmPyV VP1 (ΔC VP1) lacking the C-terminal 116 amino acid residues and compared its VLP formation efficiency and morphology to those of VLPs from wild-type VmPyV VP1 (WT VP1). WT and ΔC VP1 VLPs were similar in size, but the number of ΔC VP1 VLPs was much lower than that of WT VP1 VLPs in VP1-expressing HEK293T cells. These results suggest that the length of VP1 is unrelated to virion morphology; however, the C-terminal region of VmPyV VP1 affects the efficiency of its VLP formation.
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影响因子：
4.8
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