Rake J B, Quiñones M A, Faller D V
Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts.
J Biol Chem. 1991 Mar 15;266(8):5348-52.
The function of ras protein and its relationship to growth-factor mediated signal transduction remain unclear. The demonstration that the expression of transforming ras (v-ras) can block the stimulation of growth-related gene expression and cell division mediated by the platelet-derived growth factor (PDGF) may provide a model for the functional interation of ras with growth factor receptors. In the current studies, we have demonstrated that this blockade by v-ras of PDGF-BB signal transduction occurs very early in signal transduction, at the level of PDGF receptor autophosphorylation. Although the expression of PDGF receptor as detected by Western blot with anti-PDGF receptor antibody was not diminished in v-ras-transformed murine Balb/c 3T3 fibroblasts, the autophosphorylation of PDGF receptor in response to ligand (recombinant PDGF-BB homodimer) stimulation was profoundly suppressed. This same phenomenon of v-ras-mediated PDGF receptor autophosphorylation inhibition was also demonstrated in normal rat kidney fibroblasts. Further, factor(s) present in v-ras-expressing fibroblasts found in the membrane fractions of these cells can dominantly inhibit the autophosphorylation of the PDGF receptor obtained from normal fibroblasts. These findings suggest a role for ras in one of the earliest steps of the signal transduction pathway.
ras蛋白的功能及其与生长因子介导的信号转导之间的关系仍不清楚。转化型ras(v-ras)的表达可阻断血小板衍生生长因子(PDGF)介导的生长相关基因表达和细胞分裂的刺激,这一发现可能为ras与生长因子受体的功能相互作用提供一个模型。在当前的研究中,我们已经证明v-ras对PDGF-BB信号转导的这种阻断发生在信号转导的非常早期阶段,即在PDGF受体自身磷酸化水平。虽然用抗PDGF受体抗体进行蛋白质印迹检测时,v-ras转化的小鼠Balb/c 3T3成纤维细胞中检测到的PDGF受体表达没有减少,但响应配体(重组PDGF-BB同型二聚体)刺激时,PDGF受体的自身磷酸化受到了显著抑制。在正常大鼠肾成纤维细胞中也证实了v-ras介导的PDGF受体自身磷酸化抑制的相同现象。此外,在这些细胞的膜组分中发现的表达v-ras的成纤维细胞中存在的因子可以显著抑制从正常成纤维细胞获得的PDGF受体的自身磷酸化。这些发现表明ras在信号转导途径的最早步骤之一中发挥作用。