title: Regulation of the Src kinase-associated phosphoprotein 55 homologue by the protein tyrosine phosphatase PTP-PEST in the control of cell motility.
abstract: PTP-PEST is a cytosolic ubiquitous protein tyrosine phosphatase (PTP) that contains, in addition to its catalytic domain, several protein-protein interaction domains that allow it to interface with several signaling pathways. Among others, PTP-PEST is a key regulator of cellular motility and cytoskeleton dynamics. The complexity of the PTP-PEST interactome underscores the necessity to identify its interacting partners and physiological substrates in order to further understand its role in focal adhesion complex turnover and actin organization. Using a modified yeast substrate trapping two-hybrid system, we identified a cytosolic adaptor protein named Src kinase-associated phosphoprotein 55 homologue (SKAP-Hom) as a novel substrate of PTP-PEST. To confirm PTP-PEST interaction with SKAP-Hom, in vitro pull down assays were performed demonstrating that the PTP catalytic domain and Proline-rich 1 (P1) domain are respectively binding to the SKAP-Hom Y260 and Y297 residues and its SH3 domain. Subsequently, we generated and rescued SKAP-Hom-deficient mouse embryonic fibroblasts (MEFs) with WT SKAP-Hom, SKAP-Hom tyrosine mutants (Y260F, Y260F/Y297F), or SKAP-Hom SH3 domain mutant (W335K). Given the role of PTP-PEST, wound-healing and trans-well migration assays were performed using the generated lines. Indeed, SKAP-Hom-deficient MEFs showed a defect in migration compared with WT-rescued MEFs. Interestingly, the SH3 domain mutant-rescued MEFs showed an enhanced cell migration corresponding potentially with higher tyrosine phosphorylation levels of SKAP-Hom. These findings suggest a novel role of SKAP-Hom and its phosphorylation in the regulation of cellular motility. Moreover, these results open new avenues by which PTP-PEST regulates cellular migration, a hallmark of metastasis.
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada.
authors.forename: Emily
authors.initials: E
authors.lastname: Ayoub
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada.
authors.forename: Anita
authors.initials: A
authors.lastname: Hall
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada.
authors.forename: Adam M
authors.initials: AM
authors.lastname: Scott
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada.
authors.forename: Mélanie J
authors.initials: MJ
authors.lastname: Chagnon
authors.affiliation: INSERM U1029 and; Université Bordeaux, Avenue des Facultés, 33 405 Talence, France.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Miquel
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada.
authors.forename: Maxime
authors.initials: M
authors.lastname: Hallé
authors.affiliation: the Division of Molecular Neurobiology, National Institute for Basic Biology, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi 444-8787, Japan, and.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Noda
authors.affiliation: INSERM U1029 and; Université Bordeaux, Avenue des Facultés, 33 405 Talence, France.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bikfalvi
authors.affiliation: From the Goodman Cancer Research Centre and Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada,. Electronic address: michel.tremblay@mcgill.ca.
authors.forename: Michel L
authors.initials: ML
authors.lastname: Tremblay
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D018345:Mice, Knockout; D020125:Mutation, Missense; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12 / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D018909:src Homology Domains
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C116431:src kinase associated phosphoprotein 2; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12; C516739:Ptpn12 protein, mouse
keywords: Cell Migration; Phosphorylation; Protein-Protein Interactions; SH3 domains; Tyrosine Protein Phosphatase (Tyrosine Phosphatase)
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