Naruse H, Scholtissek C, Klenk H D
Virus Res. 1986 Aug;5(2-3):293-305. doi: 10.1016/0168-1702(86)90025-0.
Nine mutants of fowl plague virus with temperature-sensitive defects in the biosynthesis of the hemagglutinin have been characterized by analyzing the processing and the intracellular location of this glycoprotein in MDCK and chick embryo cells. It was found that with all of these mutants the transport of the hemagglutinin to the cell surface was impeded at the non-permissive temperature. There were differences, however, in the site of the block. With mutants tsl, ts227, ts478 and ts658 the precursor HA was not cleaved and the oligosaccharide side chains remained sensitive to endoglucosaminidase H. When the hemagglutinin was analyzed in permeabilized cells by immunofluorescence, usually only cytoplasmic labeling was seen. Immunofluorescence of non-permeabilized cells and hemadsorption revealed that the hemagglutinin did not reach the cell surface. In contrast, the hemagglutinin of mutants ts79, ts482, ts532, ts546 and ts651 was cleaved and oligosaccharides were processed to the endoglucosaminidase H-resistant form at non-permissive temperature. In permeabilized cells, the cytoplasm and juxtanuclear regions typical for the Golgi apparatus were labeled by immunofluorescence. Except for ts482, ts532 and ts546 which were leaky, hemagglutinin could not be detected at the cell surface. These observations indicate that, with the first group of mutants, hemagglutinin transport is usually arrested already in the rough endoplasmic reticulum, whereas with the second group it is inhibited at a late stage between the Golgi apparatus and the plasma membrane.
通过分析血凝素在MDCK细胞和鸡胚细胞中的加工过程及细胞内定位，对9种在血凝素生物合成方面具有温度敏感性缺陷的禽痘病毒突变体进行了表征。结果发现，在非允许温度下，所有这些突变体的血凝素向细胞表面的转运均受到阻碍。然而，阻断的位点存在差异。对于突变体tsl、ts227、ts478和ts658，前体HA未被切割，寡糖侧链对内切葡糖胺酶H仍敏感。当通过免疫荧光在通透细胞中分析血凝素时，通常只能看到细胞质标记。非通透细胞的免疫荧光和血细胞吸附显示血凝素未到达细胞表面。相比之下，突变体ts79、ts482、ts532、ts546和ts651的血凝素在非允许温度下被切割，寡糖被加工成对内切葡糖胺酶H有抗性的形式。在通透细胞中，免疫荧光标记了高尔基体典型的细胞质和近核区域。除了ts482、ts532和ts546有渗漏外，在细胞表面未检测到血凝素。这些观察结果表明，对于第一组突变体，血凝素转运通常在粗面内质网中就已停滞，而对于第二组突变体，其在高尔基体和质膜之间的后期阶段受到抑制。
