title: Wiskott-Aldrich syndrome protein is associated with the adapter protein Grb2 and the epidermal growth factor receptor in living cells.
abstract: Src homology domains [i.e., Src homology domain 2 (SH2) and Src homology domain 3 (SH3)] play a critical role in linking receptor tyrosine kinases to downstream signaling networks. A well-defined function of the SH3-SH2-SH3 adapter Grb2 is to link receptor tyrosine kinases, such as the epidermal growth factor receptor (EGFR), to the p21ras-signaling pathway. Grb2 has also been implicated to play a role in growth factor-regulated actin assembly and receptor endocytosis, although the underlying mechanisms remain unclear. In this study, we show that Grb2 interacts through its SH3 domains with the human Wiskott-Aldrich syndrome protein (WASp), which plays a role in regulation of the actin cytoskeleton. We find that WASp is expressed in a variety of cell types and is exclusively cytoplasmic. Although the N-terminal SH3 domain of Grb2 binds significantly stronger than the C-terminal SH3 domain to WASp, full-length Grb2 shows the strongest binding. Both phosphorylation of WASp and its interaction with Grb2, as well as with another adapter protein Nck, remain constitutive in serum-starved or epidermal growth factor-stimulated cells. WASp coimmunoprecipitates with the activated EGFR after epidermal growth factor stimulation. Purified glutathione S-transferase-full-length-Grb2 fusion protein, but not the individual domains of Grb2, enhances the association of WASp with the EGFR, suggesting that Grb2 mediates the association of WASp with EGFR. This study suggests that Grb2 translocates WASp from the cytoplasm to the plasma membrane and the Grb2-WASp complex may play a role in linking receptor tyrosine kinases to the actin cytoskeleton.
authors.affiliation: Ben May Institute for Cancer Research, University of Chicago, Illinois 60637, USA.
authors.forename: H Y
authors.initials: HY
authors.lastname: She
authors.forename: S
authors.initials: S
authors.lastname: Rockow
authors.forename: J
authors.initials: J
authors.lastname: Tang
authors.forename: R
authors.initials: R
authors.lastname: Nishimura
authors.forename: E Y
authors.initials: EY
authors.lastname: Skolnik
authors.forename: M
authors.initials: M
authors.lastname: Chen
authors.forename: B
authors.initials: B
authors.lastname: Margolis
authors.forename: W
authors.initials: W
authors.lastname: Li
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002460:Cell Line; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D051380:GRB2 Adaptor Protein; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D007668:Kidney; D051379:Mice; D015513:Oncogene Proteins / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism* / Q000502:physiology; D014407:Tumor Cells, Cultured; D014923:Wiskott-Aldrich Syndrome / Q000235:genetics / Q000378:metabolism*; D051316:Wiskott-Aldrich Syndrome Protein; D018909:src Homology Domains
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; C495949:Grb2 protein, mouse; C078332:Nck protein; D015513:Oncogene Proteins; D011506:Proteins; C495768:Was protein, mouse; D051316:Wiskott-Aldrich Syndrome Protein; D066246:ErbB Receptors