Tani H, Nishijima M, Ushijima H, Miyamura T, Matsuura Y
Department of Virology II, National Institute of Infectious Diseases, Tokyo, 162-8640.
Virology. 2001 Jan 5;279(1):343-53. doi: 10.1006/viro.2000.0699.
Baculovirus gp64 envelope glycoprotein is a major component of the envelope of the budded virus and is involved in virus entry into the host cells by endocytosis. To investigate the cell-surface molecules important for infection of baculovirus into mammalian cells, we constructed a recombinant baculovirus, Ac64-CAluc, which has gp64 and luciferase genes under the polyhedrin and the CAG promoter, respectively. For controls, we constructed recombinant viruses possessing vesicular stomatitis virus (VSV) G protein, mouse hepatitis virus (MHV) S protein, or green fluorescent protein (GFP) gene under the polyhedrin promoter and the luciferase gene under the CAG promoter (AcVSVG-CAluc, AcMHVS-CAluc, and AcGFP-CAluc). Treatment of HepG2 cells with phospholipase C markedly reduced the reporter gene expression by Ac64-CAluc or AcVSVG-CAluc in a dose-dependent manner, whereas AcMHVS-CAluc was shown to be resistant to the treatment. Inhibition with purified lipids and susceptibility to the mutant CHO hamster cell lines deficient in phospholipids synthesis suggest that the interaction of gp64 and phospholipids on the cell surface might play an important role in baculovirus infection into mammalian cells.
杆状病毒糖蛋白64（gp64）包膜糖蛋白是出芽病毒包膜的主要成分，参与病毒通过内吞作用进入宿主细胞的过程。为了研究对杆状病毒感染哺乳动物细胞起重要作用的细胞表面分子，我们构建了一种重组杆状病毒Ac64-CAluc，其在多角体蛋白启动子和CAG启动子下分别具有gp64和荧光素酶基因。作为对照，我们构建了在多角体蛋白启动子下具有水疱性口炎病毒（VSV）G蛋白、小鼠肝炎病毒（MHV）S蛋白或绿色荧光蛋白（GFP）基因，以及在CAG启动子下具有荧光素酶基因的重组病毒（AcVSVG-CAluc、AcMHVS-CAluc和AcGFP-CAluc）。用磷脂酶C处理HepG2细胞，可显著降低Ac64-CAluc或AcVSVG-CAluc的报告基因表达，且呈剂量依赖性，而AcMHVS-CAluc对该处理具有抗性。用纯化脂质进行抑制以及对磷脂合成缺陷的突变型中国仓鼠卵巢（CHO）细胞系的敏感性表明，细胞表面gp64与磷脂的相互作用可能在杆状病毒感染哺乳动物细胞中起重要作用。
