Markland W, Smith A E, Roberts B L
Integrated Genetics, Framingham, Massachusetts 01701.
Mol Cell Biol. 1987 Dec;7(12):4255-65. doi: 10.1128/mcb.7.12.4255-4265.1987.
An in vitro nuclear translocation system is described in which isolated rat liver nuclei were incubated in a defined buffered medium containing radiolabeled or fluorescently labeled exogenous proteins. The nuclei were rapidly recovered, extracted, and analyzed for the presence of associated radiolabeled or fluorescently labeled proteins. The isolated nuclei exhibited the same specificity for protein uptake as seen previously in vivo, accumulating simian virus 40 wild-type large-T antigen and p53 while excluding a cytoplasmic variant of large-T antigen (d10) and bovine serum albumin. The rapid nuclear accumulation of wild-type large-T antigen was shown to be selective and dependent upon the recognition of a wild-type nuclear location signal, ATP and temperature dependent, and unidirectional. Taken together, the data suggest that in our in vitro system the nuclear translocation of wild-type large-T antigen exhibits some of the characteristics of an active transport process.
本文描述了一种体外核转运系统，其中将分离的大鼠肝细胞核在含有放射性标记或荧光标记的外源蛋白的特定缓冲培养基中孵育。迅速回收细胞核，进行提取，并分析相关放射性标记或荧光标记蛋白的存在情况。分离的细胞核对蛋白质摄取表现出与先前在体内观察到的相同特异性，积累猿猴病毒40野生型大T抗原和p53，同时排除大T抗原的细胞质变体（d10）和牛血清白蛋白。野生型大T抗原的快速核积累被证明具有选择性，依赖于对野生型核定位信号的识别，依赖于ATP和温度，并且是单向的。综上所述，数据表明在我们的体外系统中，野生型大T抗原的核转运表现出一些主动运输过程的特征。
