title: p130Cas mediates the transforming properties of the anaplastic lymphoma kinase.
abstract: Translocations of the anaplastic lymphoma kinase (ALK) gene have been described in anaplastic large-cell lymphomas (ALCLs) and in stromal tumors. The most frequent translocation, t(2;5), generates the fusion protein nucleophosmin (NPM)-ALK with intrinsic tyrosine kinase activity. Along with transformation, NPM-ALK induces morphologic changes in fibroblasts and lymphoid cells, suggesting a direct role of ALK in cell shaping. In this study, we used a mass-spectrometry-based proteomic approach to search for proteins involved in cytoskeleton remodeling and identified p130Cas (p130 Crk-associated substrate) as a novel interactor of NPM-ALK. In 293 cells and in fibroblasts as well as in human ALK-positive lymphoma cell lines, NPM-ALK was able to bind p130Cas and to induce its phosphorylation. Both of the effects were dependent on ALK kinase activity and on the adaptor protein growth factor receptor-bound protein 2 (Grb2), since no binding or phosphorylation was found with the kinase-dead mutant NPM-ALK(K210R) or in the presence of a Grb2 dominant-negative protein. Phosphorylation of p130Cas by NPM-ALK was partially independent from Src (tyrosine kinase pp60c-src) kinase activity, as it was still detectable in Syf-/- cells. Finally, p130Cas-/- (also known as Bcar1-/-) fibroblasts expressing NPM-ALK showed impaired actin filament depolymerization and were no longer transformed compared with wild-type cells, indicating an essential role of p130Cas activation in ALK-mediated transformation.
authors.affiliation: Center for Experimental Research and Medical Studies and Department of Biomedical Sciences and Human Oncology, University of Turin, Turin, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Ambrogio
authors.forename: Claudia
authors.initials: C
authors.lastname: Voena
authors.forename: Andrea D
authors.initials: AD
authors.lastname: Manazza
authors.forename: Roberto
authors.initials: R
authors.lastname: Piva
authors.forename: Ludovica
authors.initials: L
authors.lastname: Riera
authors.forename: Laura
authors.initials: L
authors.lastname: Barberis
authors.forename: Carlotta
authors.initials: C
authors.lastname: Costa
authors.forename: Guido
authors.initials: G
authors.lastname: Tarone
authors.forename: Paola
authors.initials: P
authors.lastname: Defilippi
authors.forename: Emilio
authors.initials: E
authors.lastname: Hirsch
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Boeri Erba
authors.forename: Shabaz
authors.initials: S
authors.lastname: Mohammed
authors.forename: Ole N
authors.initials: ON
authors.lastname: Jensen
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palestro
authors.forename: Giorgio
authors.initials: G
authors.lastname: Inghirami
authors.forename: Roberto
authors.initials: R
authors.lastname: Chiarle
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D000818:Animals; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D002471:Cell Transformation, Neoplastic / Q000378:metabolism*; D050719:Crk-Associated Substrate Protein / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D016403:Lymphoma, Large B-Cell, Diffuse / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D015514:Oncogene Proteins, Fusion / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases; D014407:Tumor Cells, Cultured
chemical_list: D050719:Crk-Associated Substrate Protein; D015514:Oncogene Proteins, Fusion; C000626173:ALK protein, human; C000626174:Alk protein, mouse; D000077548:Anaplastic Lymphoma Kinase; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases