Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1420-4. doi: 10.1073/pnas.0910782107. Epub 2010 Jan 4.
AB-type toxins, like other bacterial toxins, are notably opportunistic molecules. They rely on target cell receptors to reach the appropriate location within the target cell where translocation of their enzymatic subunits occurs. The anthrax toxin, however, times its own uptake, suggesting that toxin binding triggers specific signaling events. Here we show that the anthrax toxin triggers tyrosine phosphorylation of its own receptors, capillary morphogenesis gene 2 and tumor endothelial marker 8, which are not endowed with intrinsic kinase activity. This is required for efficient toxin uptake because endocytosis of the mutant receptor lacking the cytoplasmic tyrosine residues is strongly delayed. Phosphorylation of the receptors was dependent on src-like kinases, which where activated upon toxin binding. Importantly, src-dependent phosphorylation of the receptor was required for its subsequent ubiquitination, which in turn was required for clathrin-mediated endocytosis. Consistently, we found that uptake of the anthrax toxin and processing of the lethal factor substrate MEK1 are inhibited by silencing of src and fyn, as well as in src and fyn knockout cells.
AB 型毒素与其他细菌毒素一样,是一种明显的机会主义分子。它们依赖靶细胞受体到达靶细胞内的适当位置,在那里发生它们的酶亚基的易位。然而,炭疽毒素会自行摄取,这表明毒素结合会触发特定的信号事件。在这里,我们表明炭疽毒素会引发其自身受体(毛细血管形态发生基因 2 和肿瘤内皮标记 8)的酪氨酸磷酸化,而这些受体本身并不具有内在的激酶活性。这是有效摄取毒素所必需的,因为缺乏细胞质酪氨酸残基的突变受体的内吞作用被强烈延迟。受体的磷酸化依赖于 src 样激酶,这些激酶在毒素结合时被激活。重要的是,受体的 src 依赖性磷酸化是其随后泛素化所必需的,而泛素化又是网格蛋白介导的内吞作用所必需的。一致地,我们发现炭疽毒素的摄取和致死因子底物 MEK1 的加工被沉默 src 和 fyn 以及 src 和 fyn 敲除细胞所抑制。