The SH2/SH3 adaptor Grb4 transduces B-ephrin reverse signals.
Bidirectional signals mediated by membrane-anchored ephrins and Eph receptor tyrosine kinases have important functions in cell-cell recognition events, including those that occur during axon pathfinding and hindbrain segmentation. The reverse signal that is transduced into B-ephrin-expressing cells is thought to involve tyrosine phosphorylation of the signal's short, conserved carboxy-terminal cytoplasmic domain. The Src-homology-2 (SH2) domain proteins that associate with activated tyrosine-phosphorylated B-subclass ephrins have not been identified, nor has a defined cellular response to reverse signals been described. Here we show that the SH2/SH3 domain adaptor protein Grb4 binds to the cytoplasmic domain of B ephrins in a phosphotyrosine-dependent manner. In response to B-ephrin reverse signalling, cells increase FAK catalytic activity, redistribute paxillin, lose focal adhesions, round up, and disassemble F-actin-containing stress fibres. These cellular responses can be blocked in a dominant-negative fashion by expression of the isolated Grb4 SH2 domain. The Grb4 SH3 domains bind a unique set of other proteins that are implicated in cytoskeletal regulation, including the Cbl-associated protein (CAP/ponsin), the Abl-interacting protein-1 (Abi-1), dynamin, PAK1, hnRNPK and axin. These data provide a biochemical pathway whereby cytoskeletal regulators are recruited to Eph-ephrin bidirectional signalling complexes.
Pubmed ID: 11557983 RIS Download
Adaptor Proteins, Signal Transducing | Amino Acid Sequence | Animals | Axin Protein | Cell Line | Cell Membrane | Cytoskeletal Proteins | Cytoskeleton | Ephrin-B1 | Glutathione Transferase | Humans | Membrane Proteins | Mice | Microfilament Proteins | Molecular Sequence Data | Neurons | Oncogene Proteins | Phosphotyrosine | Protein Binding | Proteins | Receptor Protein-Tyrosine Kinases | Receptor, EphB4 | Receptors, Eph Family | Recombinant Fusion Proteins | Repressor Proteins | Signal Transduction | Two-Hybrid System Techniques | src Homology Domains