DISSECTION OF THE GOLGI-COMPLEX .1. MONENSIN INHIBITS THE TRANSPORT OF VIRAL MEMBRANE-PROTEINS FROM MEDIAL TO TRANS GOLGI CISTERNAE IN BABY HAMSTER-KIDNEY CELLS INFECTED WITH SEMLIKI FOREST VIRUS
DISSECTION OF THE GOLGI-COMPLEX .1. MONENSIN INHIBITS THE TRANSPORT OF VIRAL MEMBRANE-PROTEINS FROM MEDIAL TO TRANS GOLGI CISTERNAE IN BABY HAMSTER-KIDNEY CELLS INFECTED WITH SEMLIKI FOREST VIRUS
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DOI：
10.1083/jcb.96.3.835
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发表时间：
1983-01-01
影响因子：
7.8
通讯作者：
WARREN, G
中科院分区：
文献类型：
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作者：
GRIFFITHS, G;QUINN, P;WARREN, G
用塞姆利基森林病毒（SFV）感染幼仓鼠肾（BHK）细胞，2小时后，用10 μ M莫能菌素处理4小时。高尔基体中的4 ~ 6个扁平池中的每一个都变得肿胀并与其他池分离。病毒膜蛋白的细胞内运输几乎被完全抑制，但它们的合成继续进行，并在莫能菌素阻断前在肿胀的高尔基池中积累。这些池结合大量的病毒核衣壳，很容易与其他肿胀的池，如封闭后的那些区分开来。这些细胞内的capture-binding膜（ICBM）没有染色的内质网（ER）（葡萄糖-6-磷酸酶）或反式高尔基池（硫胺素焦磷酸酶，酸性磷酸酶）的细胞化学标记，但蓖麻凝集素I（RCA）在薄，冷冻切片标记。由于这种凝集素只标记位于堆栈中间和反侧的高尔基池，因此ICBM显然来自堆栈中间的高尔基池，称为内侧池。病毒膜蛋白的整体运动似乎是从顺式到反式高尔基池（见上文参考文献），因此莫能菌素会阻止从内侧池到反式池的运动。它还阻断了与病毒膜蛋白结合的高甘露糖寡糖的修剪及其向复合寡糖的转化。这些功能可能存在于反高尔基池。显然，脂肪酸在顺式或中间池中与病毒膜蛋白共价连接。高尔基体堆叠显然可以分为3个功能不同的隔室，每个隔室包括1或2个池。病毒膜蛋白在离开ER后，将全部按顺式隔室到中间隔室再到反式隔室的顺序传递。
Baby hamster kidney (BHK) cells were infected with Semliki Forest virus (SFV) and, 2 h later, were treated for 4 h with 10 .mu.M monensin. Each of the 4 to 6 flattened cisternae in the Golgi stack became swollen and separated from the others. Intracellular transport of the viral membrane proteins was almost completely inhibited, but their synthesis continued and they accumulated in the swollen Golgi cisternae before the monensin block. These cisternae bound large numbers of viral nucleocapsids and were easily distinguished from other swollen cisternae such as those after the block. These intracellular capsid-binding membranes (ICBM) were not stained by cytochemical markers for endoplasmic reticulum (ER) (glucose-6-phosphatase) or trans Golgi cisternae (thiamine pyrophosphatase, acid phosphatase) but were labeled by Ricinus communis agglutin in I (RCA) in thin, frozen sections. Since this lectin labels only Golgi cisternae in the middle and on the trans side of the stack ICBM, apparently are derived from Golgi cisternae in the middle of the stack, which is termed medial cisternae. The overall movement of viral membrane proteins appears to be from cis to trans Golgi cisternae (see reference above), so monensin would block movement from medial to the trans cisternae. It also blocked the trimming of the high-mannose oligosaccharides bound to the viral membrane proteins and their conversion to complex oligosaccharides. These functions presumably reside in trans Golgi cisternae. Evidently, fatty acids are covalently attached to the viral membrane proteins in the cis or medial cisternae. The Golgi stack apparently can be divided into 3 functionally distinct compartments, each comprising 1 or 2 cisternae. The viral membrane proteins, after leaving the ER, would all pass in sequence from the cis to the medial to the trans compartment.