Characterization of the budding compartment of mouse hepatitis virus: evidence that transport from the RER to the Golgi complex requires only one vesicular transport step.
Characterization of the budding compartment of mouse hepatitis virus: evidence that transport from the RER to the Golgi complex requires only one vesicular transport step.
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小鼠肝炎病毒的萌芽室的表征：从RER到高尔基体配合物的运输的证据仅需要一个囊泡运输步骤。
DOI：
10.1083/jcb.124.1.55
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发表时间：
1994-01
影响因子：
7.8
通讯作者：
Griffiths, G
中科院分区：
文献类型：
--
作者：
Krijnse-Locker, J;Ericsson, M;Rottier, P J;Griffiths, G
小鼠肝炎冠状病毒（MHV）芽形成多形性膜结构，具有内质网和高尔基复合体之间的中间室的预期特征。在这里，我们使用链球菌溶血素 O (SLO) 透化技术更详细地描述了小鼠 L 细胞中 MHV 出芽室的特征，这使我们能够更好地可视化 ER-高尔基体边界的膜结构。 MHV 出芽室与粗内质网以及高尔基体堆叠一侧的池元件共享膜连续性。它还标记有 p58 和 rab2（中间室的两个标记）以及 PDI（通常被认为是粗 ER 的标记）。出芽室的膜，以及出芽病毒粒子本身，但不是粗糙的内质网，用 N-乙酰半乳糖胺 (GalNAc) 特异性凝集素 Helix pomatia 标记。当用鸟苷 5'-(3-O-硫代)三磷酸 (GTP gamma S) 处理 SLO 透化细胞时，出芽室积累了大量含有 β-cop 的芽和囊泡轮廓。进行了补充生化实验以确定新合成的仅包含 O-连接寡糖的 M 蛋白是否需要囊泡运输，以首先获得 GalNAc，然后获得高尔基体修饰的半乳糖和唾液酸。体内研究和使用 SLO 透化细胞的结果表明，虽然 GalNAc 添加发生在阻断囊泡运输的条件下，但细胞质和 ATP 都是 M 蛋白寡糖获得高尔基体修饰的先决条件。总的来说，我们的数据表明，从粗面内质网到高尔基复合体的运输只需要一个囊泡运输步骤，并且中间室是内质网的一个特殊区域，延伸到高尔基体堆叠顺侧的第一个池。
Mouse hepatitis coronavirus (MHV) buds into pleomorphic membrane structures with features expected of the intermediate compartment between the ER and the Golgi complex. Here, we characterize the MHV budding compartment in more detail in mouse L cells using streptolysin O (SLO) permeabilization which allowed us to better visualize the membrane structures at the ER-Golgi boundary. The MHV budding compartment shares membrane continuities with the rough ER as well as with cisternal elements on one side of the Golgi stack. It also labeled with p58 and rab2, two markers of the intermediate compartment, and with PDI, usually considered to be a marker of the rough ER. The membranes of the budding compartment, as well as the budding virions themselves, but not the rough ER, labeled with the N-acetyl- galactosamine (GalNAc)-specific lectin Helix pomatia. When the SLO- permeabilized cells were treated with guanosine 5'-(3-O- thio)triphosphate (GTP gamma S), the budding compartment accumulated a large number of beta-cop-containing buds and vesicular profiles. Complementary biochemical experiments were carried out to determine whether vesicular transport was required for the newly synthesized M protein, that contains only O-linked oligosaccharides, to acquire first, GalNAc and second, the Golgi modifications galactose and sialic acid. The results from both in vivo studies and from the use of SLO- permeabilized cells showed that, while GalNAc addition occurred under conditions which block vesicular transport, both cytosol and ATP were prerequisites for the M protein oligosaccharides to acquire Golgi modifications. Collectively, our data argue that transport from the rough ER to the Golgi complex requires only one vesicular transport step and that the intermediate compartment is a specialized domain of the endoplasmatic reticulum that extends to the first cisterna on the cis side of the Golgi stack.