Ceresa Brian P, Bahr Steven J
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, 73190, USA.
J Biol Chem. 2006 Jan 13;281(2):1099-106. doi: 10.1074/jbc.M504175200. Epub 2005 Nov 10.
The epidermal growth factor receptor (EGFR) is a member of the receptor tyrosine kinase family. Ligand (epidermal growth factor or EGF) binding to the EGFR results in the coordinated activation and integration of biochemical signaling events to mediate cell growth, migration, and differentiation. One mechanism the cell utilizes to orchestrate these events is ligand-mediated endocytosis through the canonical clathrin-mediated endocytic pathway. Identification of proteins that regulate the intracellular movement of the EGF.EGFR complex is an important first step in dissecting how specificity of EGFR signaling is conferred. We examined the role of the small molecular weight guanine nucleotide-binding protein (G-protein) rab7 as a regulator of the distal stages of the endocytic pathway. Through the transient expression of activating and inactivating mutants of rab7 in HeLa cells, we have determined that rab7 activity directly correlates with the rate of radiolabeled EGF and EGFR degradation. Furthermore, when inhibitory mutants of rab7 are expressed, the internalized EGF.EGFR complex accumulates in high-density endosomes that are characteristic of the late endocytic pathway. Thus, we conclude that rab7 regulates the endocytic trafficking of the EGF.EGFR complex by regulating its lysosomal degradation.
表皮生长因子受体(EGFR)是受体酪氨酸激酶家族的一员。配体(表皮生长因子或EGF)与EGFR结合会导致生化信号事件的协同激活和整合,从而介导细胞生长、迁移和分化。细胞用来协调这些事件的一种机制是通过经典的网格蛋白介导的内吞途径进行配体介导的内吞作用。鉴定调节EGF.EGFR复合物细胞内运动的蛋白质是剖析EGFR信号特异性如何赋予的重要第一步。我们研究了小分子鸟嘌呤核苷酸结合蛋白(G蛋白)rab7作为内吞途径远端阶段调节剂的作用。通过在HeLa细胞中瞬时表达rab7的激活和失活突变体,我们确定rab7活性与放射性标记的EGF和EGFR降解速率直接相关。此外,当表达rab7的抑制性突变体时,内化的EGF.EGFR复合物会积聚在晚期内吞途径特有的高密度内体中。因此,我们得出结论,rab7通过调节其溶酶体降解来调节EGF.EGFR复合物的内吞运输。