Phytopathology. 1998 Feb;88(2):98-104. doi: 10.1094/PHYTO.1998.88.2.98.
ABSTRACT Interactions between viral and cellular membrane fusion proteins mediate virus penetration of cells for many arthropod-borne viruses. Electron microscope observations and circumstantial evidence indicate insect acquisition of tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae) is receptor mediated, and TSWV membrane glycoproteins (GP1 and GP2) serve as virus attachment proteins. The tospoviruses are plant-infecting members of the family Bunyaviridae and are transmitted by several thrips species, including Frankliniella occidentalis. Gel overlay assays and immunolabeling were used to investigate the putative role of TSWV GPs as viral attachment proteins and deter mine whether a corresponding cellular receptor may be present in F. occidentalis. A single band in the 50-kDa region was detected with murine monoclonal antibodies (MAbs) to the TSWV-GPs when isolated TSWV or TSWV-GPs were used to overlay separated thrips proteins. This band was not detected when blots were probed with antibody to the non-structural protein encoded by the small RNA of TSWV or the TSWV nucleocapsid protein, nor were proteins from nonvector insects labeled. Anti-idiotype antibodies prepared to murine MAbs against GP1 or GP2 specifically labeled a single band at 50 kDa in Western blots and the plasmalemma of larval thrips midguts. These results support the putative role of the TSWV GPs as viral attachment proteins and identified potential cellular receptor(s) in thrips.
摘要 病毒和细胞膜融合蛋白之间的相互作用介导了许多节肢动物传播病毒对细胞的穿透。电子显微镜观察和间接证据表明,昆虫获得番茄斑萎病毒(TSWV)(属 Tospovirus, Bunyaviridae 科)是受体介导的,TSWV 膜糖蛋白(GP1 和 GP2)作为病毒附着蛋白。Tospoviruses 是 Bunyaviridae 科中感染植物的成员,由包括西方花蓟马(Frankliniella occidentalis)在内的几种蓟马物种传播。凝胶覆盖测定和免疫标记用于研究 TSWV GPs 作为病毒附着蛋白的可能作用,并确定 F. occidentalis 中是否存在相应的细胞受体。当使用分离的 TSWV 或 TSWV-GPs 覆盖分离的蓟马蛋白时,用针对 TSWV-GPs 的鼠单克隆抗体(MAb)检测到 50-kDa 区域的单个带。当用针对 TSWV 小 RNA 编码的非结构蛋白或 TSWV 核衣壳蛋白的抗体探测印迹时,该带未被检测到,非载体昆虫的蛋白质也未被标记。针对 GP1 或 GP2 的鼠 MAb 制备的抗独特型抗体在 Western blot 中特异性标记 50 kDa 处的单个带,以及幼虫蓟马中肠的质膜。这些结果支持 TSWV GPs 作为病毒附着蛋白的假设作用,并在蓟马中鉴定出潜在的细胞受体。