A SINGLE MUTATION IN CHINESE-HAMSTER OVARY CELLS IMPAIRS BOTH GOLGI AND ENDOSOMAL FUNCTIONS
A SINGLE MUTATION IN CHINESE-HAMSTER OVARY CELLS IMPAIRS BOTH GOLGI AND ENDOSOMAL FUNCTIONS
复制标题
DOI：
10.1083/jcb.99.4.1296
复制
发表时间：
1984-01-01
影响因子：
7.8
通讯作者：
MELLMAN, I
中科院分区：
文献类型：
--
作者：
ROBBINS, AR;OLIVER, C;MELLMAN, I
中国仓鼠卵巢细胞突变体，DTG 1-5-4，选择受体介导的内吞作用的多效性缺陷，通过先前描述的方法。DTG 1-5-4表现出增加的抗性modeccin，假单胞菌毒素，白喉毒素，辛德毕斯病毒和水泡性口炎病毒，以及通过甘露糖6-磷酸受体的摄取减少。从DTG 1-5-4中分离的白细胞介素-葡聚糖标记的内体在体外缺乏ATP依赖性酸化。在DTG 1-5-4的回复突变体和DTG 1 - 5 - 4与DTF 1-5-1的杂交体中，内吞和内体酸化都恢复，DTF 1-5-1是另一种表现出降低的ATP依赖性内体酸化的内吞突变体。DTG 1-5-4和DTF 1-5-1在辛德毕斯病毒感染的2个阶段均被阻断：在感染病毒的低多重性下，抗性反映了病毒渗透到细胞质中的阻断，但在感染病毒的高多重性下，阻断是在病毒释放中。与内吞作用一样，在DTG 1-5-4和DTG 1-5- 4x的回复突变体中，辛德毕斯病毒的释放增加。DTF 1-5-1混合体。DTG 1-5-4释放的病毒减少与辛毕斯糖蛋白成熟的一些高尔基体相关步骤中的缺陷相关;前体pE 2的蛋白水解加工、半乳糖基化和转运至细胞表面均受到抑制。Sindbis糖蛋白的甘露糖基化、岩藻糖基化和酰化以及水泡性口炎病毒和细胞糖蛋白的半乳糖基化在突变体和亲本中分别发生了相似的程度。辛德毕斯感染的DTG 1-5-4的EM检查显示与邻近高尔基体的池结合的核衣壳显著积累;在这些池的一些腔中观察到病毒体。内吞作用和高尔基体相关的病毒成熟步骤的改变是由单一的遗传损伤引起的，这表明这些过程依赖于一种共同的生化机制。内吞途径和分泌途径可能共享参与离子转运的共同组分。
A Chinese hamster ovary cell mutant, DTG 1-5-4, was selected for pleiotropic defects in receptor-mediated endocytosis by methods previously described. DTG 1-5-4 exhibited increased resistance to modeccin, Pseudomonas toxin, diphtheria toxin, Sindbis virus and vesicular stomatitis virus, as well as decreased uptake via the mannose 6-phosphate receptor. Fluorescein-dextran-labeled endosomes isolated from DTG 1-5-4 were deficient in ATP-dependent acidication in vitro. Endocytosis and endosome acidification were both restored in revertants of DTG 1-5-4 and in hybrids of DTG 1-5-4 with DTF 1-5-1, another endocytosis mutant exhibiting decreased ATP-dependent endosome acidification. Both DTG 1-5-4 and DTF 1-5-1 were blocked at 2 stages of infection with Sindbis virus: at low multiplicities of infecting virus, resistance reflected a block in viral penetration into the cytoplasm, but at higher multiplicities of infection the block was in virus release. Like endocytosis, release of Sindbis virus was increased in revertants of DTG 1-5-4 and in DTG 1-5-4 .times. DTF 1-5-1 hybrids. Decreased release of virus from DTG 1-5-4 correlated with defects in some of the Golgi apparatus-associated steps of Sindbis glycoprotein maturation; proteolytic processing of the precursor pE2, galactosylation and transport to the cell surface all were inhibited. Mannosylation, fucosylation and acylation of the Sindbis glycoproteins, and galactosylation of vesicular stomatitis virus and cellular glycoprotein occurred to similar respective extents in mutant and parent. EM examination of Sindbis-infected DTG 1-5-4 showed a remarkable accumulation of nucleocapsids bound to cisternae adjacent to the Golgi apparatus; virions were observed in the lumina of some of these cisternae. That the alterations in both endocytosis and Golgi-associated steps of viral maturation result from a single genetic lesion indicates that these processes are dependent on a common biochemical mechanism. Endocytic and secretory pathways may share a common component involved in ion transport.