Neurite extension and protein tyrosine phosphorylation elicited by inducible expression of the v-src oncogene in a PC12 cell line.
Neurite extension and protein tyrosine phosphorylation elicited by inducible expression of the v-src oncogene in a PC12 cell line.
复制标题
PC12 细胞系中 v-src 癌基因的诱导表达引起神经突延伸和蛋白质酪氨酸磷酸化。
DOI：
10.1016/0014-4827(91)90393-9
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发表时间：
1991
影响因子：
3.7
通讯作者：
Maness,PF
中科院分区：
文献类型：
--
作者：
Cox,ME;Maness,PF
在PC 12嗜铬细胞瘤细胞系（PC 12-B 9）中研究了酪氨酸特异性蛋白激酶活性，所述细胞系通过稳定转染在小鼠金属硫蛋白酶I基因启动子的转录控制下编码活化酪氨酸激酶的诱导型v-src基因（pp 60 v-src）而产生。用Cd ~（2+）和Zn ~（2+）诱导PC_（12-B 9）细胞表达pp 60 v-src，可使PC_（12-B 9）细胞可逆性分化为神经元样细胞。pp 60 v-src基因表达促进PC 12-B 9细胞形态分化，并具有明显的一级动力学，与NGF诱导的神经突起生长速率相同，v-src基因表达以累加方式促进NGF诱导的神经突起生长速率。诱导pp 60 v-srcalone组成性增加磷酸酪氨酸修饰的蛋白质（130-120，90，83，65，60/59，36 kDa）的水平，通过免疫印迹检测与磷酸酪氨酸抗体。NGF处理的PC 12-B 9细胞瞬时增加了不同的磷酸酪氨酸修饰的蛋白质（108，46，42 kDa），以及共同的底物，包括一个59 kDa的蛋白质与α-微管蛋白共迁移的水平。磷酸酪氨酸修饰的蛋白质在v-src和NGF诱导的PC 12-B 9细胞中没有协同增加。v-src基因表达对神经突起生长和磷酸化的非协同效应表明pp 60 v-src通过与NGF诱导的细胞内信号传导途径很大程度上不同的细胞内信号传导途径诱导PC 12细胞分化。
Tyrosine-specific protein kinase activity in neuronal differentiation was studied in a PC12 pheochromocytoma cell line (PC12-B9) produced by stable transfection with an inducible v-srcgene encoding an activated tyrosine kinase (pp60v-src) under the transcriptional control of the mouse metallothionine I gene promoter. Induction of pp60v-srcexpression with Cd2+and Zn2+resulted in the reversible differentiation of PC12-B9 cells into neuron-like cells. pp60v-srcelicited morphological differentiation with apparent first order kinetics at the same rate as NGF-directed neurite outgrowth in PC12-B9 cells, v-srcgene expression enhanced the rate of NGF-directed neurite extension in an additive manner. Induction of pp60v-srcalone constitutively increased the levels of phosphotyrosine-modified proteins (130-120, 90, 83, 65, 60/59, 36 kDa) detected by immunoblotting with phosphotyrosine antibodies. NGF treatment of PC12-B9 cells transiently increased the levels of distinct phosphotyrosine-modified proteins (108, 46, 42 kDa), as well as common substrates, including a 59-kDa protein that comigrated with α-tubulin. Phosphotyrosine-modified proteins were not synergistically increased in PC12-B9 cells induced for both v-srcand NGF. The nonsynergistic effects of v-srcgene expression on neurite outgrowth and phosphorylation suggest that pp60v-srcinduces PC12 cell differentiation by an intracellular signaling pathway that is largely distinct from that induced by NGF.