Shi Z Q, Lu W, Feng G S
Department of Biochemistry and Molecular Biology, Walther Oncology Center, Indiana University School of Medicine and Walther Cancer Institute, Indianapolis, Indiana 46202-5121, USA.
J Biol Chem. 1998 Feb 27;273(9):4904-8. doi: 10.1074/jbc.273.9.4904.
Shp-2 is a widely expressed cytoplasmic tyrosine phosphatase with two SH2 domains. A targeted mutant allele of the Shp-2 gene with a deletion of 65 amino acids in the NH2-terminal SH2 domain was created that leads to embryonic lethality at mid-gestation in homozygous mutant mice. To define the Shp-2 function in cell signaling, we have established mutant fibroblast cell lines, and have examined the effect of the Shp-2 mutation on extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) mitogen-activated protein (MAP) kinase pathways. Insulin-like growth factor (IGF)-I-induced ERK activation was completely abolished, while ERK activity upon platelet-derived growth factor and epidermal growth factor stimulation was significantly reduced and shortened in mutant cells. Stimulation of ERK by phorbol 12-myristate 13-acetate was not affected in mutant cells, but the phorbol 12-myristate 13-acetate-induced ERK activity decayed much faster compared with that in wild-type cells. In contrast, JNK activation upon heat shock was significantly enhanced in Shp-2 mutant cells. Based on these results, we conclude that Shp-2 plays differential positive regulatory roles in various mitogenic signaling pathways leading to ERK activation, and that Shp-2 is a negative effector in JNK activation by cellular stress. This is the first evidence that a tyrosine phosphatase has opposite effects in mediating the activation of ERK and JNK MAP kinases.
Shp-2是一种广泛表达的细胞质酪氨酸磷酸酶,具有两个SH2结构域。构建了一个Shp-2基因的靶向突变等位基因,其NH2末端SH2结构域缺失65个氨基酸,导致纯合突变小鼠在妊娠中期胚胎致死。为了确定Shp-2在细胞信号传导中的功能,我们建立了突变成纤维细胞系,并研究了Shp-2突变对细胞外信号调节激酶(ERK)和c-Jun NH2末端激酶(JNK)丝裂原活化蛋白(MAP)激酶途径的影响。胰岛素样生长因子(IGF)-I诱导的ERK激活完全被消除,而血小板衍生生长因子和表皮生长因子刺激后的ERK活性在突变细胞中显著降低且持续时间缩短。佛波酯12-肉豆蔻酸酯13-乙酸酯对ERK的刺激在突变细胞中不受影响,但与野生型细胞相比,佛波酯12-肉豆蔻酸酯13-乙酸酯诱导的ERK活性衰减得更快。相反,在热休克后,Shp-2突变细胞中的JNK激活显著增强。基于这些结果,我们得出结论,Shp-2在导致ERK激活的各种促有丝分裂信号通路中发挥不同的正向调节作用,并且Shp-2是细胞应激激活JNK的负效应物。这是酪氨酸磷酸酶在介导ERK和JNK MAP激酶激活中具有相反作用的首个证据。