Tashiro Kyoko, Tsunematsu Takumi, Okubo Hiroko, Ohta Takeshi, Sano Etsuko, Yamauchi Emiko, Taniguchi Hisaaki, Konishi Hiroaki
Division of Disease Proteomics, Institute for Enzyme Research, the University of Tokushima, Tokushima 770-8503, Japan.
J Biol Chem. 2009 Jul 24;284(30):20206-14. doi: 10.1074/jbc.M109.021139. Epub 2009 Jun 9.
Adaptor proteins for the various growth factor receptors play a crucial role in signal transduction through tyrosine phosphorylation. Several candidates for adaptor proteins with potential effects on the epidermal growth factor (EGF) receptor-mediated signaling pathway have been identified by recent phosphoproteomic studies. Here, we focus on a novel protein, GAREM (Grb2-associated and regulator of Erk/MAPK) as a downstream molecule of the EGF receptor. GAREM is phosphorylated at tyrosine 105 and 453 after EGF stimulation. Grb2 was identified as its binding partner, and the proline-rich motifs of GAREM are recognized by the N- and C-terminal SH3 domains of Grb2. In addition, the tyrosine phosphorylations of GAREM are necessary for its binding to Grb2. Because the amino acid sequence surrounding tyrosine 453 is similar to the immunoreceptor tyrosine-based inhibitory motif, Shp2, a positive regulator of Erk, binds to GAREM in this phosphorylation-dependent manner. Consequently, Erk activation in response to EGF stimulation is regulated by the expression of GAREM in COS-7 and HeLa cells, which occurs independent of the presence of other binding proteins, such as Gab1 and SOS, to the activated EGF receptor. Furthermore, the expression of GAREM has an effect on the transformation activity of cultured cells. Together, these findings suggest that GAREM plays a key role in the ligand-mediated signaling pathway of the EGF receptor and the tumorigenesis of cells.
各种生长因子受体的衔接蛋白在通过酪氨酸磷酸化的信号转导中起关键作用。近期的磷酸化蛋白质组学研究已鉴定出几种对表皮生长因子(EGF)受体介导的信号通路可能有影响的衔接蛋白候选物。在此,我们聚焦于一种新型蛋白质GAREM(Grb2相关且为Erk/丝裂原活化蛋白激酶的调节因子),它是EGF受体的下游分子。EGF刺激后,GAREM在酪氨酸105和453处发生磷酸化。Grb2被鉴定为其结合伴侣,GAREM富含脯氨酸的基序被Grb2的N端和C端SH3结构域识别。此外,GAREM的酪氨酸磷酸化对于其与Grb2的结合是必需的。由于酪氨酸453周围的氨基酸序列与免疫受体酪氨酸抑制基序相似,Erk的正向调节因子Shp2以这种磷酸化依赖的方式与GAREM结合。因此,在COS - 7和HeLa细胞中,对EGF刺激的Erk激活受GAREM表达的调节,这种调节独立于其他与活化的EGF受体结合的蛋白(如Gab1和SOS)的存在。此外,GAREM的表达对培养细胞的转化活性有影响。总之,这些发现表明GAREM在EGF受体的配体介导信号通路和细胞肿瘤发生中起关键作用。