Volchkov V E, Volchkova V A, Slenczka W, Klenk H D, Feldmann H
Institut für Virologie, Philipps-Universität Marburg, Germany.
Virology. 1998 May 25;245(1):110-9. doi: 10.1006/viro.1998.9143.
Maturation and release of the Ebola virus glycoprotein GP were studied in cells infected with either Ebola or recombinant vaccinia viruses. Significant amounts of GP were found in the culture medium in nonvirion forms. The major form represented the large subunit GP1 that was shed after release of its disulfide linkage to the smaller transmembrane subunit GP2. The minor form were intact GP1,2 complexes incorporated into virosomes. Vector-expressed GP formed spikes morphologically indistinguishable from spikes on virus particles, indicating that spike assembly is independent of other viral proteins. Analysis of a truncation mutant revealed an early and almost complete release of GP1,2 molecules, showing that membrane anchoring is mediated by the carboxy-terminal hydrophobic domain of GP2. We have also compared wild-type virus which requires transcriptional editing for synthesis of full-length GP with a variant that does not depend on editing. Both viruses released comparable amounts of GP1, but the variant expressed only minute amounts of the small, soluble GP which is the expression product of nonedited mRNA species of the GP gene. The abundant shedding of soluble GP1 may play an important role in the immunopathology of Ebola hemorrhagic fever in experimentally and naturally infected hosts.
在感染埃博拉病毒或重组痘苗病毒的细胞中研究了埃博拉病毒糖蛋白GP的成熟和释放。在培养基中发现了大量非病毒形式的GP。主要形式是大亚基GP1,其与较小的跨膜亚基GP2的二硫键断裂后脱落。次要形式是完整的GP1,2复合物,整合到病毒体中。载体表达的GP形成的刺突在形态上与病毒颗粒上的刺突无法区分,表明刺突组装独立于其他病毒蛋白。对截短突变体的分析显示GP1,2分子早期几乎完全释放,表明膜锚定由GP2的羧基末端疏水结构域介导。我们还比较了需要转录编辑以合成全长GP的野生型病毒和不依赖编辑的变体。两种病毒释放的GP1量相当,但该变体仅表达微量的小的可溶性GP,其是GP基因未编辑mRNA种类的表达产物。可溶性GP1的大量脱落可能在实验性和自然感染宿主中埃博拉出血热的免疫病理学中起重要作用。