Li Y, Basilico C, Mansukhani A
Department of Microbiology and Kaplan Cancer Center, New York University School of Medicine, New York 10016.
Mol Cell Biol. 1994 Nov;14(11):7660-9. doi: 10.1128/mcb.14.11.7660-7669.1994.
Ligand-induced dimerization and transphosphorylation are thought to be important events by which receptor tyrosine kinases generate cellular signals. We have investigated the ability of signalling-defective, truncated fibroblast growth factor (FGF) receptors (FGFR-1 and FGFR-2) to block the FGF response in cells that express both types of endogenous FGF receptors. When these dominant negative receptors are expressed in NIH 3T3 cells transformed by the secreted FGF-4, the transformed properties of the cells can be reverted to various degrees, with better reversion phenotype correlating with higher levels of truncated receptor expression. Furthermore, truncated FGFR-2 is significantly more efficient at producing reversion than FGFR-1, indicating that FGF-4 preferentially utilizes the FGFR-2 signalling pathway. NIH 3T3 clones expressing these truncated receptors are more resistant to FGF-induced mitogenesis and also exhibit reduced tyrosine phosphorylation upon treatment with FGF. The block in FGF-signalling, however, can be overcome by the addition of excess growth factor. The truncated receptors have binding affinities that are four- to eightfold lower than those of wild-type receptors, as measured by Scatchard analysis. We also observed a partial specificity in the responses of truncated-receptor-expressing clones to FGF-2 or FGF-4. Our results suggest that the block to signal transduction produced by kinase-negative FGF receptors is achieved through a combination of dominant negative effects and competition for growth factor binding with functional receptors.
配体诱导的二聚化和转磷酸化被认为是受体酪氨酸激酶产生细胞信号的重要事件。我们研究了信号缺陷型截短成纤维细胞生长因子(FGF)受体(FGFR-1和FGFR-2)在表达两种内源性FGF受体的细胞中阻断FGF反应的能力。当这些显性负性受体在由分泌型FGF-4转化的NIH 3T3细胞中表达时,细胞的转化特性可在不同程度上恢复,恢复表型越好与截短受体表达水平越高相关。此外,截短的FGFR-2在产生恢复方面比FGFR-1明显更有效,表明FGF-4优先利用FGFR-2信号通路。表达这些截短受体的NIH 3T3克隆对FGF诱导的有丝分裂更具抗性,在用FGF处理时酪氨酸磷酸化也减少。然而,通过添加过量生长因子可以克服FGF信号传导的阻断。通过Scatchard分析测量,截短受体的结合亲和力比野生型受体低4至8倍。我们还观察到表达截短受体的克隆对FGF-2或FGF-4的反应存在部分特异性。我们的结果表明,激酶阴性FGF受体产生的信号转导阻断是通过显性负性效应和与功能性受体竞争生长因子结合的组合来实现的。