title: Tensin1 requires protein phosphatase-1alpha in addition to RhoGAP DLC-1 to control cell polarization, migration, and invasion.
abstract: Tensin is a family of multidomain scaffold proteins that bind the cytoplasmic tail of beta-integrins and localize to adhesions that anchor stress fibers in cells. Tensin expression is suppressed in cancer, especially metastatic cancer. The N-terminal domain of tensin1 associates with protein phosphatase-1alpha (PP1alpha) and mediates PP1alpha localization to adhesions. Here, we show F302A mutation in a KVXF motif of tensin1 abrogates binding to PP1alpha. The SH2 domain in tensin family member c-ten requires R474 to bind a RhoGAP called DLC-1 (deleted in liver cancer). We mutated the corresponding residue in tensin1, R1488A, and showed this reduces association with DLC-1. Unexpectedly, tensin1 F302A also had reduced association with DLC-1. Expression of tensin1 F302A or R1488A showed similar dominant phenotypes, with reduced cell polarization, lowered MLC20 phosphorylation and reduced levels of RhoA(GTP) compared with cells expressing tensin1 WT. However, migration and invasion of metastatic MDA MB 231 breast cancer cells were differentially affected by tensin1 mutated at F302A or R1488A. Cancer cells stably expressing F302A tensin1 showed increased migration and invasion compared with cells stably expressing either R1488A tensin1 or WT tensin1. This suggests that PP1alpha bound to tensin1 has additional effects in reducing migration and invasion that are not mediated through DLC-1. Our results show the importance of PP1alpha binding to tensin1 for the regulation of cell polarization, migration, and invasion.
authors.affiliation: Center for Cell Signaling and Department of Microbiology, University of Virginia School of Medicine, Charlottesville, Virginia 22908, USA.
authors.forename: Emily H
authors.initials: EH
authors.lastname: Hall
authors.forename: Abbi E
authors.initials: AE
authors.lastname: Daugherty
authors.forename: Colin K
authors.initials: CK
authors.lastname: Choi
authors.forename: Alan F
authors.initials: AF
authors.lastname: Horwitz
authors.forename: David L
authors.initials: DL
authors.lastname: Brautigan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D016764:Cell Polarity; D048430:Cell Shape; D004789:Enzyme Activation; D020690:GTPase-Activating Proteins; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness / Q000503:physiopathology; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology; D017354:Point Mutation; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000070876:Tensins; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: C112352:DLC1 protein, human; D020690:GTPase-Activating Proteins; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; C000603369:TNS1 protein, human; D000070876:Tensins; D025521:Tumor Suppressor Proteins; D054645:Protein Phosphatase 1; D020742:rhoA GTP-Binding Protein