title: Src kinase determines the dynamic exchange of the docking protein NEDD9 (neural precursor cell expressed developmentally down-regulated gene 9) at focal adhesions.
abstract: Dynamic exchange of molecules between the cytoplasm and integrin-based focal adhesions provides a rapid response system for modulating cell adhesion. Increased residency time of molecules that regulate adhesion turnover contributes to adhesion stability, ultimately determining migration speed across two-dimensional surfaces. In the present study we test the role of Src kinase in regulating dynamic exchange of the focal adhesion protein NEDD9/HEF1/Cas-L. Using either chemical inhibition or fibroblasts genetically null for Src together with fluorescence recovery after photobleaching (FRAP), we find that Src significantly reduces NEDD9 exchange at focal adhesions. Analysis of NEDD9 mutant constructs with the two major Src-interacting domains disabled revealed the greatest effects were due to the NEDD9 SH2 binding domain. This correlated with a significant change in two-dimensional migratory speed. Given the emerging role of NEDD9 as a regulator of focal adhesion stability, the time of NEDD9 association at the focal adhesions is key in modulating rates of migration and invasion. Our study suggests that Src kinase activity determines NEDD9 exchange at focal adhesions and may similarly modulate other focal adhesion-targeted Src substrates to regulate cell migration.
authors: From the Children's Cancer Research Unit, Kids Research Institute at The Children's Hospital at Westmead, Westmead 2145, New South Wales and Discipline of Paediatrics and Child Health, University of Sydney, New South Wales 2006, Australia.
authors: Peta
authors: P
authors: Bradbury
authors: From the Children's Cancer Research Unit, Kids Research Institute at The Children's Hospital at Westmead, Westmead 2145, New South Wales and.
authors: Cuc T
authors: CT
authors: Bach
authors: From the Children's Cancer Research Unit, Kids Research Institute at The Children's Hospital at Westmead, Westmead 2145, New South Wales and.
authors: Andre
authors: A
authors: Paul
authors: From the Children's Cancer Research Unit, Kids Research Institute at The Children's Hospital at Westmead, Westmead 2145, New South Wales and Discipline of Paediatrics and Child Health, University of Sydney, New South Wales 2006, Australia geraldine.oneill@health.nsw.gov.au.
authors: Geraldine M
authors: GM
authors: O'Neill
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D022001:Focal Adhesions / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D007700:Kinetics; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D009154:Mutation; D013997:Time Factors; D018909:src Homology Domains / Q000235:genetics; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C505592:NEDD9 protein, mouse; D049452:Green Fluorescent Proteins; D019061:src-Family Kinases
keywords: Cell Adhesion; Cell Biology; Cell Migration; Fluorescence; PTK2 Protein-tyrosine Kinase 2 (PTK2) (Focal Adhesion Kinase) (FAK)