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© The Rockefeller University Press, 0021-9525/2000//1419 $5.00
The Journal of Cell Biology, Volume 149, Number 7, , 2000 1419-1432

Original Article

Coupling of Gab1 to C-Met, Grb2, and Shp2 Mediates Biological Responses



Ute Schaepera, Niels H. Gehringa, Klaus P. Fuchsb, Martin Sachsa, Bettina Kempkesb, and Walter Birchmeiera

a Max Delbrück Center for Molecular Medicine, 13092 Berlin, Germany
b Institute for Clinical Molecular Biology and Tumor Genetics, GSF Research Center for Environment and Health, 81377 Munich, Germany
Max Delbrück Center for Molecular Medicine Berlin-Buch, Robert Rössle Str. 10, 13092 Berlin, Germany.49 30 9406 265649 30 9406 3810

uschaep{at}mdc-berlin.de

Gab1 is a substrate of the receptor tyrosine kinase c-Met and involved in c-Met–specific branching morphogenesis. It associates directly with c-Met via the c-Met–binding domain, which is not related to known phosphotyrosine-binding domains. In addition, Gab1 is engaged in a constitutive complex with the adaptor protein Grb2. We have now mapped the c-Met and Grb2 interaction sites using reverse yeast two-hybrid technology. The c-Met–binding site is localized to a 13–amino acid region unique to Gab1. Insertion of this site into the Gab1-related protein p97/Gab2 was sufficient to confer c-Met–binding activity. Association with Grb2 was mapped to two sites: a classical SH3-binding site (PXXP) and a novel Grb2 SH3 consensus-binding motif (PX(V/I)(D/N)RXXKP). To detect phosphorylation-dependent interactions of Gab1 with downstream substrates, we developed a modified yeast two-hybrid assay and identified PI(3)K, Shc, Shp2, and CRKL as interaction partners of Gab1. In a trk-met-Gab1–specific branching morphogenesis assay, association of Gab1 with Shp2, but not PI(3)K, CRKL, or Shc was essential to induce a biological response in MDCK cells. Overexpression of a Gab1 mutant deficient in Shp2 interaction could also block HGF/SF-induced activation of the MAPK pathway, suggesting that Shp2 is critical for c-Met/Gab1-specific signaling.

Key Words: Gab1 • c-Met–binding site • morphogenesis • Shp2 • reverse yeast two-hybrid analysis

© 2000 The Rockefeller University Press

Abbreviations used in this paper: CSW, Corkscrew; DOS, daughter of Sevenless; Gab1, Grb2-associated binder 1; HA, hemagglutinin; IRS, insulin receptor substrates; MBS, c-Met–binding site; PI(3)K, phosphatidylinositol 3-kinase; PLC-{gamma}, phospholipase C-{gamma}; SH, Src homology.


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