title: Dissecting motility signaling through activation of specific Src-effector complexes.
abstract: We describe an approach to selectively activate a kinase in a specific protein complex or at a specific subcellular location within living cells and within minutes. This reveals the effects of specific kinase pathways without time for genetic compensation. The new technique, dubbed rapamycin-regulated targeted activation of pathways (RapRTAP), was used to dissect the role of Src kinase interactions with FAK and p130Cas in cell motility and morphodynamics. The overall effects of Src activation on cell morphology and adhesion dynamics were first quantified, without restricting effector access. Subsets of Src-induced behaviors were then attributed to specific interactions between Src and the two downstream proteins. Activation of Src in the cytoplasm versus at the cell membrane also produced distinct phenotypes. The conserved nature of the kinase site modified for RapRTAP indicates that the technique can be applied to many kinases.
authors.affiliation: 1] Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA. [2].
authors.forename: Andrei V
authors.initials: AV
authors.lastname: Karginov
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Denis
authors.initials: D
authors.lastname: Tsygankov
authors.affiliation: Department of Biomedical Engineering, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Berginski
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Pei-Hsuan
authors.initials: PH
authors.lastname: Chu
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Evan D
authors.initials: ED
authors.lastname: Trudeau
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Jason J
authors.initials: JJ
authors.lastname: Yi
authors.affiliation: Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Shawn
authors.identifier: 0000000282514552
authors.initials: S
authors.lastname: Gomez
authors.affiliation: Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Elston
authors.affiliation: 1] Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA. [2] Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Hahn
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D002465:Cell Movement / Q000187:drug effects* / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D050719:Crk-Associated Substrate Protein / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology / Q000648:ultrastructure; D051417:Focal Adhesion Kinase 1 / Q000235:genetics / Q000378:metabolism; D016391:Genes, src / Q000187:drug effects*; D006801:Humans; D008856:Microscopy, Fluorescence; D010641:Phenotype; D011494:Protein Kinases / Q000187:drug effects*; D011554:Pseudopodia / Q000187:drug effects / Q000648:ultrastructure; D015398:Signal Transduction / Q000187:drug effects*; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C493841:BCAR1 protein, human; D050719:Crk-Associated Substrate Protein; D011494:Protein Kinases; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D022021:Tacrolimus Binding Proteins