Nucleocytoplasmic transport is enhanced concomitant with nuclear accumulation of epidermal growth factor (EGF) binding activity in both 3T3-1 and EGF receptor reconstituted NR-6 fibroblasts.
Nucleocytoplasmic transport is enhanced concomitant with nuclear accumulation of epidermal growth factor (EGF) binding activity in both 3T3-1 and EGF receptor reconstituted NR-6 fibroblasts.
复制标题
在3T3-1和EGF受体重建的NR-6成纤维细胞中，核质转运与表皮生长因子（EGF）结合活性的核积累增强。
DOI：
10.1083/jcb.110.3.559
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发表时间：
1990-03
影响因子：
7.8
通讯作者：
Schindler, M
中科院分区：
文献类型：
--
作者：
Jiang, L W;Schindler, M
在EGF无反应的突变型成纤维细胞系(3T3-NR6)和由来自大鼠肝细胞膜部分的活性EGF受体重组的同一细胞系中，对荧光标记的大分子的核质转运进行了测量。重组3T3-NR6成纤维细胞中外源性掺入的EGF受体的免疫定位研究表明，主要定位于细胞内。EGF受体构建体还显示了EGF刺激的[~3H]胸腺嘧啶核苷的掺入，为外源提供的EGF受体功能整合到重组的成纤维细胞中提供了生化证据。氯喹和亮肽素处理的细胞中125I-EGF的增加可能是对受体功能整合的额外支持。突变和重组细胞的125I-EGF结合和跨核大分子转运的测量，与从这些细胞分离的核的这种测量相结合，提供了与生长因子/核信号机制一致的数据，该机制依赖于核从质膜获得EGF结合活性。
Measurements of nucleocytoplasmic transport of fluorescent-labeled macromolecules were performed in both an EGF-nonresponsive mutant fibroblast line (3T3-NR6) and in the same cell line reconstituted with active EGF receptors derived from rat hepatic membrane fraction. Immunolocalization studies of exogenously incorporated EGF receptors in reconstituted 3T3-NR6 fibroblasts demonstrated predominantly intracellular localization. The EGF receptor constructs also showed EGF- stimulated incorporation of [3H]thymidine, providing biochemical evidence for functional integration of the exogenously supplied EGF receptors into the reconstituted fibroblasts. Additional support for the functional incorporation of receptor may be inferred from the enhanced cellular accumulation of 125I-EGF in cells treated with chloroquine and leupeptin. 125I-EGF binding and transnuclear macromolecular transport measurements in mutant and reconstituted cells, in conjunction with such measurements on nuclei isolated from these cells, provide data consistent with a growth factor/nuclear signaling mechanism dependent on the nuclear acquisition of EGF binding activity from the plasma membrane.