A TEMPERATURE-SENSITIVE CHINESE-HAMSTER OVARY CELL MUTANT PLEIOTROPICALLY DEFECTIVE IN PROTEIN EXPORT
A TEMPERATURE-SENSITIVE CHINESE-HAMSTER OVARY CELL MUTANT PLEIOTROPICALLY DEFECTIVE IN PROTEIN EXPORT
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DOI：
10.1016/0167-4889(85)90195-8
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发表时间：
1985-01-01
期刊：
影响因子：
--
通讯作者：
AKAMATSU, Y
中科院分区：
文献类型：
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作者：
NAKANO, A;NISHIJIMA, M;AKAMATSU, Y
本文提出了一种新的利用白喉毒素筛选蛋白质分泌缺陷型哺乳动物细胞突变株的方法，并设计了一种利用硝酸纤维素膜筛选蛋白质分泌缺陷型突变株的新方法。通过这些程序的组合，中国仓鼠卵巢细胞的温度敏感性突变克隆，其在蛋白质输出中显示多效性缺陷。该突变体命名为DS 28 -6，对生长温度敏感。在非允许温度下，一系列蛋白质的分泌被显著抑制。这些蛋白质似乎在非允许温度下在细胞内正常合成和积累，并在降低至允许温度时分泌。当该突变体感染水泡性口炎病毒时，G蛋白糖蛋白的寡糖加工在非允许温度下被阻止在内切糖基化酶-H-敏感阶段。这种突变体的病变似乎是在内质网或顺高尔基体或两者兼而有之。
A new selection procedure for mammalian cell mutants defective in protein export by the use of diphtheria toxin was developed, and a new screening method was devised for defective protein secretion using nitrocellulose membranes. By the combination of these procedures, a temperature-sensitive mutant clone of Chinese hamster ovary cells which shows a pleiotropic defect in protein export was isolated. This mutant, designated DS28-6, is temperature-sensitive for growth. Secretion of a series of proteins is markedly inhibited at the non-permissive temperature. These proteins seem to be normally synthesized and accumulated within the cell at the non-permissive temperature and secreted upon shift down to the permissive temperature. When this mutant is infected with vesicular stomatitis virus, oligosaccharide processing of G-protein glycoprotein is arrested at an endoglycosidase-H-sensitive stage at the non-permissive temperature. The lesion of this mutant appears to be in the endoplasmic reticulum or the cis Golgi or both.