Hyaluronan promotes CD44v3-Vav2 interaction with Grb2-p185(HER2) and induces Rac1 and Ras signaling during ovarian tumor cell migration and growth

• Published in: The Journal of biological chemistry 2001-12, Vol.276 (52), p.48679-48692
• Main Authors: Bourguignon, L Y, Zhu, H, Zhou, B, Diedrich, F, Singleton, P A, Hung, M C
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing; Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - metabolism; Cell Division - physiology; Cell Movement - physiology; Female; GRB2 Adaptor Protein; Guanine Nucleotide Exchange Factors - metabolism; Guanosine Triphosphate - analogs & derivatives; Guanosine Triphosphate - metabolism; Humans; Hyaluronan Receptors - genetics; Hyaluronan Receptors - metabolism; Hyaluronic Acid - metabolism; Models, Biological; Oncogene Proteins - genetics; Oncogene Proteins - metabolism; Ovarian Neoplasms - metabolism; Ovarian Neoplasms - pathology; Protein Binding; Proteins - genetics; Proteins - metabolism; Proto-Oncogene Proteins c-vav; rac1 GTP-Binding Protein - metabolism; Radioligand Assay; ras GTPase-Activating Proteins - metabolism; Receptor, ErbB-2 - genetics; Receptor, ErbB-2 - metabolism; Signal Transduction - physiology; src Homology Domains - genetics; Tumor Cells, Cultured
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M106759200

In this study we initially examined the interaction between CD44v3 (a hyaluronan (HA) receptor) and Vav2 (a guanine nucleotide exchange factor) in human ovarian tumor cells (SK-OV-3.ipl cell line). Immunological data indicate that both CD44v3 and Vav2 are expressed in SK-OV-3.ipl cells and that these two proteins are physically linked as a complex in vivo. By using recombinant fragments of Vav2 and in vitro binding assays, we have detected a specific binding interaction between the SH3-SH2-SH3 domain of Vav2 and the cytoplasmic domain of CD44. In addition, we have observed that the binding of HA to CD44v3 activates Vav2-mediated Rac1 signaling leading to ovarian tumor cell migration. Further analyses indicate that the adaptor molecule, growth factor receptor-bound protein 2 (Grb2) that is bound to p185(HER2) (an oncogene product), is also associated with the CD44v3-Vav2 complex. HA binding to SK-OV-3.ipl cells promotes recruitment of both Grb2 and p185(HER2) to the CD44v3-Vav2 complex leading to Ras activation and ovarian tumor cell growth. In order to determine the role of Grb2 in CD44v3 signaling, we have transfected SK-OV-3.ipl cells with Grb2 mutant cDNAs (e.g. Delta N-Grb2 that has a deletion in the amino-terminal SH3 domain or Delta C-Grb2 that has a deletion in the carboxyl-terminal SH3 domain). Our results clearly indicate that the SH3 domain deletion mutants of Grb2 (i.e. the Delta N-Grb2 (and to a lesser extent the Delta C-Grb2) mutant) not only block their association with p185(HER2) but also significantly impair their binding to the CD44v3-Vav2 complex and inhibit HA/CD44v3-induced ovarian tumor cell behaviors. Taken together, these findings strongly suggest that the interaction of CD44v3-Vav2 with Grb2-p185(HER2) plays an important role in the co-activation of both Rac1 and Ras signaling that is required for HA-mediated human ovarian tumor progression.