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Papers In Press, published online ahead of print December 9, 2002
Cold Spring Harbor Laboratory, Cold Spring harbor, NY 11724
Corresponding Author: tonks{at}cshl.org
The receptor protein-tyrosine phosphatase DEP-1 (CD148/PTP-
J. Biol. Chem, 10.1074/jbc.M210656200
Submitted on October 17, 2002
Revised on December 3, 2002
Accepted on December 9, 2002
"Hepatocyte growth factor receptor tyrosine kinase met is a substrate of the receptor protein-tyrosine phosphatase DEP-1"
) has been implicated in the regulation of cell growth, differentiation and transformation and most recently has been identified as a potential tumor suppressor gene mutated in colon, lung and breast cancers. We have generated constructs comprising the cytoplasmic segment of DEP-1 fused to the maltose binding protein to identify potential substrates and thereby suggest a physiological function for DEP-1. We have shown that the substrate-trapping mutant form of DEP-1 interacted with a small subset of tyrosine-phosphorylated proteins from lysates of the human breast tumor cell lines MDA-MB-231, T-47D and T-47D/Met and have identified the hepatocyte growth factor/scatter factor (HGF/SF) receptor Met, the adapter protein Gab1 and the junctional component p120 catenin as potential substrates. Following ligand stimulation, phosphorylation of specific tyrosyl residues in Met induces mitogenic, motogenic and morphogenic responses. When co-expressed in 293 cells, the full-length substrate trapping mutant form of DEP-1 formed a stable complex with the chimeric receptor CSF.MET and wild type DEP-1 dephosphorylated CSF.MET. Furthermore, we observed that DEP-1 preferentially dephosphorylated a Gab1 binding site (Tyr1349) and a C-terminal tyrosine implicated in morphogenesis (Tyr1365), whereas tyrosine residues in the activation loop of Met (Tyr1230, Tyr1234, Tyr1235) were not preferred targets of the PTP. The ability of DEP-1 preferentially to dephosphorylate particular tyrosine residues that are required for Met-induced signaling suggests that DEP-1 may function in controlling the specificity of signals induced by this PTK, rather than as a simple off-switch to counteract PTK activity.
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