School of Pathology and Laboratory Medicine, University of Western Australia, Crawley 6009,Western Australia.
J Biol Chem. 2010 Apr 2;285(14):10969-81. doi: 10.1074/jbc.M109.094920. Epub 2010 Feb 4.
The ability of thymocytes to assess T cell receptor (TCR) signaling strength and initiate the appropriate downstream response is crucial for determining their fate. We have previously shown that a c-Cbl RING finger mutant knock-in mouse, in which the E3 ubiquitin ligase activity of c-Cbl is inactivated, is highly sensitive to TCR-induced death signals that cause thymic deletion. This high intensity signal involves the enhanced tyrosine phosphorylation of the mutant c-Cbl protein promoting a marked increase in the activation of Akt. Here we show that this high intensity signal in c-Cbl RING finger mutant thymocytes also promotes the enhanced induction of two mediators of TCR-directed thymocyte apoptosis, Nur77 and the pro-apoptotic Bcl-2 family member, Bim. In contrast, a knock-in mouse harboring a mutation at Tyr-737, the site in c-Cbl that activates phosphatidylinositol 3-kinase, shows reduced TCR-mediated responses including suppression of Akt activation, a reduced induction of Nur77 and Bim, and greater resistance to thymocyte death. These findings identify tyrosine-phosphorylated c-Cbl as a critical sensor of TCR signal strength that regulates the engagement of death-promoting signals.
胸腺细胞评估 T 细胞受体 (TCR) 信号强度并启动适当的下游反应的能力对于决定其命运至关重要。我们之前曾表明,一种 c-Cbl RING 指突突变敲入小鼠,其中 c-Cbl 的 E3 泛素连接酶活性被失活,对导致胸腺细胞缺失的 TCR 诱导的死亡信号高度敏感。这种高强度信号涉及突变 c-Cbl 蛋白的酪氨酸磷酸化增强,促进 Akt 的显著激活。在这里,我们表明,c-Cbl RING 指突突变的胸腺细胞中的这种高强度信号也促进了 TCR 定向胸腺细胞凋亡的两种介质的增强诱导,Nur77 和促凋亡 Bcl-2 家族成员 Bim。相比之下,一种携带 c-Cbl 中激活磷脂酰肌醇 3-激酶的位点 Tyr-737 突变的敲入小鼠显示出减少的 TCR 介导的反应,包括 Akt 激活的抑制、Nur77 和 Bim 的诱导减少以及对胸腺细胞死亡的抗性增加。这些发现确定了酪氨酸磷酸化的 c-Cbl 作为 TCR 信号强度的关键传感器,它调节促进死亡信号的参与。