Fibroblast growth factor receptor-1-mediated endothelial cell proliferation is dependent on the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk
Fibroblast growth factor receptor-1-mediated endothelial cell proliferation is dependent on the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk
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DOI：
10.1074/jbc.274.36.25726
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发表时间：
1999-09-03
影响因子：
4.8
通讯作者：
Claesson-Welsh, L
中科院分区：
文献类型：
--
作者：
Larsson, H;Klint, P;Claesson-Welsh, L
刺激内皮细胞上表达的成纤维细胞生长因子受体-2 （FGFR-1）可导致细胞迁移和增殖。我们已经研究了Src同源性（SH） 2/SH3结构域的接头蛋白Crk在这些过程中的作用。在成纤维细胞生长因子-2刺激的内皮细胞中，Crk的瞬时酪氨酸磷酸化依赖于FGFR-1中近膜酪氨酸残基463，Crk SH2结构域通过磷酸化的Tyr-463沉淀FGFR-1，表明Crk和FGFR-1之间直接形成复合物。此外，Crk SH2和SH3结构域与Shc、C3G和Crk相关底物（Cas）形成了与配体无关的配合物。C3G和Cas的酪氨酸磷酸化增加是生长因子处理的结果。我们通过使用表达嵌合血小板衍生生长因子受体- α /FGFR-1 （α R/FR）野生型和突变型Y463F受体的细胞，研究了Crk在FGFR-1介导的细胞反应中的作用。α R/FR Y463F的激酶活性是完整的，但在突变细胞中，Crk和接头FRS-2都不再被酪氨酸磷酸化。野生型和突变型受体细胞都能有效迁移，而表达突变型α R/FR Y463F的细胞不能增殖，Erk2和Jun激酶活性在这些细胞中受到抑制。在短暂表达Crk SH2结构域突变的野生型α R/FR细胞中，Erk和Jun激酶活性以及DNA合成被减弱。我们的数据表明，Crk参与FGFR-1下游的信号复合物，传播有丝分裂信号。
Stimulation of fibroblast growth factor receptor-2 (FGFR-1) expressed on endothelial cells leads to cellular migration and proliferation. We have examined the role of the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk in these processes. Transient tyrosine phosphorylation of Crk in fibroblast growth factor-2-stimulated endothelial cells was dependent on the juxtamembrane tyrosine residue 463 in FGFR-1, and a Crk SH2 domain precipitated FGFR-1 via phosphorylated Tyr-463, indicating direct complex formation between Crk and FGFR-1. Furthermore, Crk SH2 and SH3 domains formed ligand-independent complexes with Shc, C3G, and the Crk-associated substrate (Cas). Tyrosine phosphorylation of C3G and Cas increased as a consequence of growth factor treatment, We examined the role of Crk in FGFR-1-mediated cellular responses by use of cells expressing chimeric platelet-derived growth factor receptor-alpha/FGFR-1 (alpha R/FR) wild type and mutant Y463F receptors. The kinase activity of alpha R/FR Y463F was intact, but both Crk and the adaptor FRS-2 were no longer tyrosine-phosphorylated in the mutant cells. Both wild type and mutant receptor cells migrated efficiently, whereas cells expressing the mutant alpha R/FR Y463F failed to proliferate and Erk2 and Jun kinase activities were suppressed in these cells. In wild type alpha R/FR cells transiently expressing an SH2 domain mutant of Crk, Erk and Jun kinase activities as well as DNA synthesis were attenuated. Our data indicate that Crk participates in signaling complexes downstream of FGFR-1, which propagate mitogenic signals.