Greene J J, Dieffenbach C W, Yang L C, Ts'o P O
Biochemistry. 1980 May 27;19(11):2485-91. doi: 10.1021/bi00552a029.
Human fibroblast and Syrian hamster embryo cells were induced to synthesize interferon (IF) with rIn . rCn and rIn . rCn + DEAE-dextran, respectively. Following induction, these cells synthesized IF for only a short time before entering into a repressed state and shutting off the synthesis of IF. Homologous and heterologous whole cell translational systems were developed to investigate the molecular basis for the shut-off of IF synthesis. These systems allowedd for the introduction of exogenous hamster and human IF-mRNAs into intact normal and repressed hamster and human cells via an improved CaCl2 precipitation technique. Human IF-mRNA was translated in normal human and hamster cells and in repressed hamster cells but not in repressed human cells. In contrast, the hamster IF-mRNA was translated in normal human, normal hamster, and repressed human cells but not in repressed hamster cells. These results indicate that a species-specific mechanism inhibiting translation of IF-mRNA is directly responsible for the shut-off of IF synthesis in human fibroblasts and Syrian hamster embryo cells.
分别用rIn、rCn以及rIn.rCn + 葡聚糖硫酸酯钠诱导人成纤维细胞和叙利亚仓鼠胚胎细胞合成干扰素(IF)。诱导后,这些细胞仅在短时间内合成IF,随后进入抑制状态并停止IF合成。开发了同源和异源全细胞翻译系统来研究IF合成停止的分子基础。这些系统通过改进的氯化钙沉淀技术,允许将外源性仓鼠和人IF - mRNA导入完整的正常和受抑制的仓鼠及人细胞中。人IF - mRNA可在正常人细胞、仓鼠细胞以及受抑制的仓鼠细胞中翻译,但不能在受抑制的人细胞中翻译。相反,仓鼠IF - mRNA可在正常人细胞、正常仓鼠细胞以及受抑制的人细胞中翻译,但不能在受抑制的仓鼠细胞中翻译。这些结果表明,一种抑制IF - mRNA翻译的物种特异性机制直接导致了人成纤维细胞和叙利亚仓鼠胚胎细胞中IF合成的停止。