TRANSFORMATION-SPECIFIC TYROSINE PHOSPHORYLATION OF A NOVEL CELLULAR PROTEIN IN CHICKEN-CELLS EXPRESSING ONCOGENIC VARIANTS OF THE AVIAN CELLULAR SRC GENE
TRANSFORMATION-SPECIFIC TYROSINE PHOSPHORYLATION OF A NOVEL CELLULAR PROTEIN IN CHICKEN-CELLS EXPRESSING ONCOGENIC VARIANTS OF THE AVIAN CELLULAR SRC GENE
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DOI：
10.1128/mcb.9.2.629
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发表时间：
1989-02-01
影响因子：
5.3
通讯作者：
PARSONS, JT
中科院分区：
文献类型：
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作者：
REYNOLDS, AB;ROESEL, DJ;PARSONS, JT
我们使用了肉豆蔻基化和非肉豆蔻基化的c-src变体和磷酸酪氨酸特异性抗体来重新评估酪氨酸磷酸化在pp 60 src细胞转化中的作用。用于检测酪氨酸磷酸化蛋白质的现有方法未能区分酪氨酸磷酸化的预测差异，这用磷酸酪氨酸特异性抗体和蛋白质印迹（免疫印迹）清楚地观察到。在这里，我们报告观察到的120，000-Mr的蛋白质，其酪氨酸磷酸化与诱导形态转化。p120在过度表达受调控的非致癌pp 60 c-src的细胞中未观察到，而在表达活化的致癌pp 60527 F的细胞中，p120的磷酸化大大增强。此外，p120的磷酸化不诱导表达的活化，但nonmyristylated src变体pp 602 A/527 F，这是转化缺陷。p120优先与细胞膜分配，这与转化SRC蛋白是膜结合的观察结果一致。虽然一些额外的推定的基板被确定和部分特征相对于细胞内定位，这些蛋白质的酪氨酸磷酸化没有紧密联系到转化。
We used myristylated and nonmyristylated c-src-based variants and phosphotyrosine-specific antibodies to reevaluate the role of tyrosine phosphorylation in cellular transformation by pp60src. Prior methods used to detect tyrosine-phosphorylated proteins failed to discriminate predicted differences in tyrosine phosphorylation which are clearly observed with phosphotyrosine-specific antibodies and Western blotting (immunoblotting). Here we report the observation of a 120,000-Mr protein whose phosphorylation on tyrosine correlates with the induction of morphological transformation. p120 was not observed in cells overexpressing the regulated, nononcogenic pp60c-src, whereas phosphorylation of p120 was greatly enhanced in cell expressing activated, oncogenic pp60527F. Furthermore, phosphorylation of p120 was not induced by expression of the activated but nonmyristylated src variant pp602A/527F, which is transformation defective. p120 partitioned preferentially with cellular membranes, consistent with the observation that transforming src proteins are membrane associated. Although a number of additional putative substrates were identified and partially characterized with respect to intracellular localization, tyrosine phosphorylation of these proteins was not tighly linked to transformation.