title: Galpha13 stimulates cell migration through cortactin-interacting protein Hax-1.
abstract: Galpha13, the alpha-subunit of the heterotrimeric G protein G13, has been shown to stimulate cell migration in addition to inducing oncogenic transformation. Cta, a Drosophila ortholog of G13, has been shown to be critical for cell migration leading to the ventral furrow formation in Drosophila embryos. Loss of Galpha13 has been shown to disrupt cell migration associated with angiogenesis in developing mouse embryos. Whereas these observations point to the vital role of G13-orthologs in regulating cell migration, widely across the species barrier, the mechanism by which Galpha13 couples to cytoskeleton and cell migration is largely unknown. Here we show that Galpha13 physically interacts with Hax-1, a cytoskeleton-associated, cortactin-interacting intracellular protein, and this interaction is required for Galpha13-stimulated cell migration. Hax-1 interaction is specific to Galpha13, and this interaction is more pronounced with the mutationally or functionally activated form of Galpha13 as compared with the wild-type Galpha13. Expression of Hax-1 reduces the formation of actin stress fibers and focal adhesion complexes in Galpha13-expressing NIH3T3 cells. Coexpression of Hax-1 also attenuates Galpha(13)-stimulated activity of Rho while potentiating Galpha13-stimulated activity of Rac. The presence of a quadnary complex consisting of Galpha13, Hax-1, Rac, and cortactin indicates the role of Hax-1 in tethering Galpha13 to the cytoskeletal component(s) involved in cell movement. Whereas the expression of Hax-1 potentiates Galpha13-mediated cell movement, silencing of endogenous Hax-1 with Hax-1-specific small interfering RNAs drastically reduces Galpha13-mediated cell migration. These findings, along with the observation that Hax-1 is overexpressed in metastatic tumors and tumor cell lines, suggest a novel role for the association of oncogenic Galpha13 and Hax-1 in tumor metastasis.
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.
authors.forename: V
authors.initials: V
authors.lastname: Radhika
authors.forename: Djamila
authors.initials: D
authors.lastname: Onesime
authors.forename: Ji Hee
authors.initials: JH
authors.lastname: Ha
authors.forename: N
authors.initials: N
authors.lastname: Dhanasekaran
mesh_terms: D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D019556:COS Cells; D002465:Cell Movement; D051356:Cortactin; D003599:Cytoskeleton / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000502:physiology*; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism*; D008954:Models, Biological; D009154:Mutation; D041681:NIH 3T3 Cells; D009362:Neoplasm Metastasis; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000502:physiology*; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D051356:Cortactin; C495869:Cttn protein, mouse; C104759:HAX1 protein, human; D008840:Microfilament Proteins; D011506:Proteins; D034741:RNA, Small Interfering; D044365:GTP-Binding Protein alpha Subunits, G12-G13