Transformation and pp60v-src autophosphorylation correlate with SHC-GRB2 complex formation in rat and chicken cells expressing host-range and kinase-active, transformation-defective alleles of v-src.
Transformation and pp60v-src autophosphorylation correlate with SHC-GRB2 complex formation in rat and chicken cells expressing host-range and kinase-active, transformation-defective alleles of v-src.
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在表达宿主范围和激酶活性、v-src 转化缺陷等位基因的大鼠和鸡细胞中，转化和 pp60v-src 自磷酸化与 SHC-GRB2 复合物的形成相关。
DOI：
10.1091/mbc.6.8.953
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发表时间：
1995
影响因子：
3.3
通讯作者：
WoodsIgnatoski,KM
中科院分区：
文献类型：
--
作者：
Verderame,MF;Guan,JL;WoodsIgnatoski,KM
几种被认为参与src介导转化的pp60v-src底物的生化特性在表达激酶活性的、转化缺陷的v-src等位基因(v-src-F172 Delta/Y416F)及其亲本等位基因v-src-F172 Delta的细胞中进行了检测，v-src-F172 Delta是一种宿主范围依赖的等位基因，可以将鸡细胞转化为梭形细胞，但不会转化大鼠细胞。由于pp60v-src-F172 Delta依赖于自身磷酸化的转化能力，这些等位基因为研究pp60v-src自磷酸化在调节底物相互作用中的作用提供了一个独特的机会。Pp125FAK酪氨酸磷酸化增加和pp60v-src相关的高水平磷酸肌醇-3‘激酶活性在表现出圆形、折射转化的鸡细胞中特异性地被检测到，但在转化为梭形形态的细胞中没有检测到。Pp125FAK活性增加，但不增加pp125FAK酪氨酸磷酸化，这与pp60v-src自磷酸化有关，并增加了锚定非依赖性生长。因此，pp125FAK和PI3‘K可能参与了v-src的形态转化。此外，磷酸化的SHC与接头Grb2的结合与增加大鼠和鸡细胞的锚定非依赖性生长(和自磷酸化)相关，而不依赖于诱导的形态表型。因此，可能通过SHC与Grb2的结合来调节宿主范围对转化的依赖，从而暗示SHC是src依赖转化的关键底物。
The biochemical properties of several pp60v-src substrates believed to participate in src-mediated transformation were examined in cells expressing a kinase-active, transformation-defective v-src allele (v-src-F172 delta/Y416F) and its parental allele, v-src-F172 delta, a host-range--dependent allele that transforms chicken cells to a fusiform morphology, but does not transform rat cells. Because pp60v-src-F172 delta is dependent on autophosphorylation for transforming ability, these alleles provide a unique opportunity to examine the role of pp60v-src autophosphorylation in regulating substrate interactions. Increased pp125FAK tyrosine phosphorylation and high levels of pp60v-src-associated phosphotidylinositol-3' kinase activity were detected specifically in chicken cells exhibiting round, refractile transformation but not in cells transformed to a fusiform morphology. Increased pp125FAK kinase activity, but not increased pp125FAK tyrosine-phosphorylation correlated with pp60v-src autophosphorylation and increased anchorage-independent growth. Thus, pp125FAK and PI3'K may participate in morphological transformation by v-src. Furthermore, association of phosphorylated SHC with the adapter GRB2 correlated with increased anchorage-independent growth (and autophosphorylation) in both rat and chicken cells independent of the morphological phenotype induced. Therefore, host-range dependence for transformation may be regulated through association of SHC with GRB2, thus implicating SHC as a crucial substrate for src-dependent transformation.