Eps15R and clathrin regulate EphB2‐mediated cell repulsion. (6th November 2017)

• Record Type: Journal Article
• Authors: Evergren, Emma; Cobbe, Neville; McMahon, Harvey T.
• Is Part Of: Traffic. Volume 19:Number 1(2018)
• Page Start: 44
• Page End: 57
• Publication Date: 2017-11-06
• Subjects: clathrin -- EphB2 -- Eps15 -- Eps15R -- Numb -- trans‐endocytosis; Biological transport -- Periodicals; 571.6
• DOI: 10.1111/tra.12531
• Languages: English
• ISSNs: 1398-9219

Abstract

Abstract : Expression of Eph receptors and their ligands, the ephrins, have important functions in boundary formation and morphogenesis in both adult and embryonic tissue. The EphB receptors and ephrinB ligands are transmembrane proteins that are expressed in different cells and their interaction drives cell repulsion. For cell repulsion to occur, trans‐endocytosis of the inter‐cellular receptor‐ligand EphB‐ephrinB complex is required. The molecular mechanism underlying trans‐endocytosis is poorly defined. Here we show that the process is clathrin‐ and Eps15R‐mediated using Co115 colorectal cell lines stably expressing EphB2 and ephrinB1. Cell repulsion in co‐cultures of EphB2‐ and ephrinB1‐expressing cells is significantly reduced by knockdown of Eps15R but not Eps15. A novel interaction motif in Eps15R, DPFxxLDPF, is shown to bind directly to the clathrin terminal domain in vitro. Moreover, the interaction between Eps15R and clathrin is required for EphB2‐mediated cell repulsion as shown in a rescue experiment in the EphB2 co‐culture assay where wild type Eps15R but not the clathrin‐binding mutant rescues cell repulsion. These results provide the first evidence that Eps15R together with clathrin control EphB/ephrinB trans‐endocytosis and thereby cell repulsion. Abstract : EphB receptors and their ligands are important for tissue boundary formation during development, and are also involved in mediating cancer cell invasion. This form of contact‐mediated cell‐cell repulsion is dependent on removal of EphB receptors from the cell surface. Here we investigate the role of clathrin‐mediated endocytosis in EphB2‐mediated cell repulsion. We show that it is both clathrin‐ and Eps15R‐dependent and identify a novel clathrin‐binding motif in Eps15R that is important for efficient cell‐cell repulsion.