Bodelón G, Labrada L, Martínez-Costas J, Benavente J
Departamento de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
Virology. 2001 Nov 25;290(2):181-91. doi: 10.1006/viro.2001.1159.
The avian reovirus S1 gene contains three partially overlapping, out-of-phase open reading frames (ORFs) that the highly conserved in all avian reovirus strains examined to date. The three S1 ORFs of the avian reovirus strain S1133 were individually expressed in bacterial cells, and their purified translation products used as antigens to raise specific polyclonal antibodies. With these antibodies we were able to demonstrate that all three S1 ORFs from different avian reovirus strains are translatable in infected cells. Proteins p10 and p17, which are specified by ORF1 and ORF2, respectively, are nonstructural proteins which associate with cell membranes, whereas ORF3 directs the synthesis of protein sigma C, a structural oligomeric protein responsible for cell attachment. While intracellular synthesis of protein sigma C was demonstrated a long time ago and that of protein p10 was reported recently, this is the first time that expression of the S1 ORF2 has been demonstrated experimentally. Thus, the previously reported coding capacity of the avian reovirus genome is now expanded to 14 proteins, of which ten are structural (lambda A, lambda B, lambda C, microA, microB, microBC, microBN, sigma A, sigma B, and sigma C) and four are nonstructural (microNS, sigma NS, p17, and p10). Finally, protein p10, but not p17 or sigma C, induces cell-cell fusion when transiently expressed in mammalian cells, supporting a previously published observation that the polypeptide encoded by the S1 ORF1 plays an important role in the syncytial phenotype displayed by avian reoviruses.
禽呼肠孤病毒S1基因包含三个部分重叠、相位不同的开放阅读框(ORF),这些开放阅读框在迄今为止检测的所有禽呼肠孤病毒毒株中高度保守。禽呼肠孤病毒毒株S1133的三个S1 ORF在细菌细胞中分别表达,其纯化的翻译产物用作抗原以产生特异性多克隆抗体。利用这些抗体,我们能够证明来自不同禽呼肠孤病毒毒株的所有三个S1 ORF在受感染细胞中均可翻译。分别由ORF1和ORF2指定的蛋白质p10和p17是非结构蛋白,它们与细胞膜相关联,而ORF3指导蛋白质sigma C的合成,sigma C是一种负责细胞附着的结构寡聚蛋白。虽然很久以前就证明了蛋白质sigma C的细胞内合成,并且最近报道了蛋白质p10的细胞内合成,但这是首次通过实验证明S1 ORF2的表达。因此,禽呼肠孤病毒基因组先前报道的编码能力现在扩展到14种蛋白质,其中10种是结构蛋白(lambda A、lambda B、lambda C、microA、microB、microBC、microBN、sigma A、sigma B和sigma C),4种是非结构蛋白(microNS、sigma NS、p17和p10)。最后,蛋白质p10而非p17或sigma C在哺乳动物细胞中瞬时表达时会诱导细胞-细胞融合,这支持了先前发表的观察结果,即S1 ORF1编码的多肽在禽呼肠孤病毒显示的合胞体表型中起重要作用。