Role of Grb7 targeting to focal contacts and its phosphorylation by focal adhesion kinase in regulation of cell migration.
We have previously described Grb7 association with focal adhesion kinase (FAK) and its possible roles in cell migration. In this paper, we investigated the mechanisms by which Grb7 and its association with FAK regulate cell migration. We found that deletion of the Grb7 SH2 domain eliminated partial Grb7 localization to focal contacts and its ability to stimulate cell migration. Replacement of the SH2 domain with the focal adhesion targeting sequence from FAK resulted in the focal contacts localization of the chimeric molecule and restored its activity to stimulate cell migration. We also found that Grb7 could be phosphorylated by FAK, which was dependent on the FAK kinase activity but not the presence of the Src family kinases. Cell adhesion also enhanced Grb7 phosphorylation in FAK+/+ cells but not FAK-/- cells, suggesting that Grb7 is a physiological substrate of FAK. Furthermore, both Grb7 and the chimeric molecule did not increase migration of FAK-/- cells, although the chimeric molecule was targeted to the focal contacts. Last, we showed that other Grb7 family members could not stimulate cell migration under similar experimental conditions. Together, these results demonstrate a role for Grb7 targeting to focal contacts and its phosphorylation by FAK in the regulation of cell migration.
Han
D C
DC
Cancer Biology Laboratories, Department of Molecular Medicine, Cornell University, Ithaca, New York 14853, USA.
Shen
T L
TL
Guan
J L
JL
D000818:Animals; D016466:CHO Cells; D002465:Cell Movement; D006224:Cricetinae; D051416:Focal Adhesion Protein-Tyrosine Kinases; D051382:GRB7 Adaptor Protein; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D011506:Proteins / Q000502:physiology*; D014443:Tyrosine / Q000378:metabolism; D018909:src Homology Domains
D011506:Proteins; D051382:GRB7 Adaptor Protein; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases