d-cbl binding to drk leads to dose-dependent down-regulation of egfr signaling and increases receptor-ligand endocytosisd-cbl绑定到井架导致下调egfr信号存在剂量依赖的相关性,增加receptor-ligand内吞作用.pdfVIP

D-Cbl Binding to Drk Leads to Dose-Dependent Down- Regulation of EGFR Signaling and Increases Receptor- Ligand Endocytosis 1 1,2,3 Pei-Yu Wang , Li-Mei Pai * 1 Graduate Institute of Biomedical Science, Chang Gung University, Tao-Yuan, Taiwan, 2 Department of Biochemistry, Chang Gung University, Tao-Yuan, Taiwan, 3 Chang Gung Molecular Medicine Research Center, Chang Gung University, Tao-Yuan, Taiwan Abstract Proper control of Epidermal Growth Factor Receptor (EGFR) signaling is critical for normal development and regulated cell behaviors. Abnormal EGFR signaling is associated with tumorigenic process of various cancers. Complicated feedback networks control EGFR signaling through ligand production, and internalization-mediated destruction of ligand-receptor complexes. Previously, we found that two isoforms of D-Cbl, D-CblS and D-CblL, regulate EGFR signaling through distinct mechanisms. While D-CblL plays a crucial role in dose-dependent down-regulation of EGFR signaling, D-CblS acts in normal restriction of EGFR signaling and does not display dosage effect. Here, we determined the underlying molecular mechanism, and found that Drk facilitates the dose-dependent regulation of EGFR signaling through binding to the proline-rich motif of D-CblL, PR. Furthermore, the RING finger domain of D-CblL is essential for promoting endocytosis of the ligand-receptor complex. Interestingly, a fusion protein of the two essential domains of D-CblL, RING- PR, is sufficient to down-regulate EGFR signal in a dose-dependent manner by promoting internalization of the ligand, Gurken. Besides, RING-SH2Drk, a fusion protein of the RING finger domain of D-Cbl and the SH2 domain of Drk, also effectively down-regulates EGFR signaling in Drosophila follicle cells, and suppresses the effects of constitutively activated