Identification of the Lymantria dispar nucleopolyhedrovirus envelope fusion protein provides evidence for a phylogenetic division of the Baculoviridae
Identification of the Lymantria dispar nucleopolyhedrovirus envelope fusion protein provides evidence for a phylogenetic division of the Baculoviridae
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DOI：
10.1128/jvi.74.13.6126-6131.2000
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发表时间：
2000-07-01
影响因子：
5.4
通讯作者：
Rohrmann, GF
中科院分区：
文献类型：
--
作者：
Pearson, MN;Groten, C;Rohrmann, GF
杆状病毒科许多不同成员的完整基因组序列，包括：核多角体病毒 (NPV) 和颗粒病毒 (GV)，显示它们缺乏 GP64 的同源物，GP64 是苜蓿银纹夜蛾多核衣壳 NPV (AcMNPV) 及其近亲的芽状形式的包膜融合蛋白。对其中一种病毒 Lymantria dispar MNPV (LdMNPV) 的基因组进行计算机辅助分析，揭示了单个开放阅读框 (ld130)，其产物具有膜蛋白的预测特性。使用免疫荧光和荧光显微镜对全长 ld130 基因和 ld130 增强型绿色荧光蛋白基因 (egfp) 融合产物的定位进行表征，结果表明 LD130 在细胞质膜上积累感染 LdMNPV 或转染 ld130-egfp。此外，用 ld130 或 ld130-egfp 转染或用野生型病毒感染的细胞在 pH 5 下进行膜融合，Western blot 分析表明 LD130 作为 83 kDa 蛋白存在于感染细胞中，并且还作为包含 81 和 83 kDa 条带的蛋白双联体存在于出芽病毒粒子中，衣霉素处理感染细胞产生约 72 kDa 的免疫反应条带，表明LD130 是 N-糖基化的，而 gp64 的分布似乎仅限于相对密切相关的 NPV 组，而 ld130 的同源物则存在于不同数量的 NPV 和 GV 中。这表明LD130可能是原始杆状病毒包膜融合蛋白。
The complete genome sequences of a number of diverse members of the Baculoviridae including: both nucleopolyhedroviruses (NPVs) and granuloviruses (GVs) revealed that they lack a homolog of GP64, the envelope fusion protein of the budded form of Autographa californica multinucleocapsid NPV (AcMNPV) and its close relatives. Computer-assisted analyses of the genome of one of these viruses, Lymantria dispar MNPV (LdMNPV), revealed a single open reading frame (ld130) whose product had the predicted properties of a membrane protein, Characterization of the localization of the products of the full-length ld130 gene and of an ld130-enhanced green fluorescent protein gene (egfp) fusion using both immunofluorescence and fluorescence microscopy revealed that LD130 accumulates at the plasma membranes of cells infected with LdMNPV or transfected with ld130-egfp. In addition, cells transfected with either ld130 or ld130-egfp or infected with wildtype virus undergo membrane fusion at pH 5, Western blot analyses indicate that LD130 is present in infected cells as an 83-kDa protein and is also present in budded virions as a protein doublet containing bands of 81 and 83 kDa, Tunicamycin treatment of infected cells resulted in an immunoreactive band of about 72 kDa, indicating that LD130 is N-glycosylated, Whereas the distribution of gp64 appears to be confined to a relatively closely related group of NPVs, homologs of ld130 are present in a diverse number of both NPVs and GVs. This suggests that LD130 may be the primordial baculovirus envelope fusion protein.