title: Dvl2 silencing in postdevelopmental cells results in aberrant cell membrane activity and actin disorganization.
abstract: The upstream events by which endothelial cells perceive the necessity for migration and how this signal results in coordinated movement is unknown. The synchrony underlying these events shares parallels to events occurring during the movement of tissues in embryogenesis. While Wnt signaling is an important pathway in development, components of the cascade exist in postdevelopment endothelial cells. The objective of this study was to determine whether Dishevelled, a key modulation protein in canonical and PCP-CE Wnt signaling was present in endothelium and its potential function. Western blots of cell lysates and immunolabeling studies confirmed that Dishevelled 2 (Dvl2) is an abundant phosphoprotein in endothelial cells. Dvl2 was localized within the cytoplasm of cells as either F-actin-free or F-actin-associated. The disappearance of F-actin-free Dvl2 in vesicle-like organelles and targeting of actin filaments correlated with a loss in cell motility. Gene silencing of Dishevelled by siRNA duplexes resulted in cells with aberrant membrane activity and an inability to extend lamellipodia. Underlying these abnormalities was a disorganization of the actin filament system, including loss of actin-rich densities, indistinct stress fibers and an accompanying increase in diffuse and aggregate cytoplasmic actin. This study represents the first documentation of Dvl2 in postdevelopmental endothelial cells and its possible role in cell migration via manipulation of actin filament bundles.
authors.affiliation: Department of Cell Biology, Hope Heart Institute, Seattle, Washington 98122, USA. awechezak@hopeheart.org
authors.forename: Arlene R
authors.initials: AR
authors.lastname: Wechezak
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Coan
mesh_terms: D000199:Actins / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism / Q000473:pathology; D002465:Cell Movement; D002478:Cells, Cultured; D000072261:Dishevelled Proteins; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D020868:Gene Silencing; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D010641:Phenotype; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D051153:Wnt Proteins
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D000072261:Dishevelled Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D051153:Wnt Proteins