title: Filamin A, the Arp2/3 complex, and the morphology and function of cortical actin filaments in human melanoma cells.
abstract: The Arp2/3 complex and filamin A (FLNa) branch actin filaments. To define the role of these actin-binding proteins in cellular actin architecture, we compared the morphology of FLNa-deficient human melanoma (M2) cells and three stable derivatives of these cells expressing normal FLNa concentrations. All the cell lines contain similar amounts of the Arp2/3 complex. Serum addition causes serum-starved M2 cells to extend flat protrusions transiently; thereafter, the protrusions turn into spherical blebs and the cells do not crawl. The short-lived lamellae of M2 cells contain a dense mat of long actin filaments in contrast to a more three-dimensional orthogonal network of shorter actin filaments in lamellae of identically treated FLNa-expressing cells capable of translational locomotion. FLNa-specific antibodies localize throughout the leading lamellae of these cells at junctions between orthogonally intersecting actin filaments. Arp2/3 complex-specific antibodies stain diffusely and label a few, although not the same, actin filament overlap sites as FLNa antibody. We conclude that FLNa is essential in cells that express it for stabilizing orthogonal actin networks suitable for locomotion. Contrary to some proposals, Arp2/3 complex-mediated branching of actin alone is insufficient for establishing an orthogonal actin organization or maintaining mechanical stability at the leading edge.
authors.affiliation: Hematology Division, Brigham and Women's Hospital, Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Flanagan
authors.forename: J
authors.initials: J
authors.lastname: Chou
authors.forename: H
authors.initials: H
authors.lastname: Falet
authors.forename: R
authors.initials: R
authors.lastname: Neujahr
authors.forename: J H
authors.initials: JH
authors.lastname: Hartwig
authors.forename: T P
authors.initials: TP
authors.lastname: Stossel
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology; D003285:Contractile Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D064448:Filamins; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008545:Melanoma; D008840:Microfilament Proteins / Q000378:metabolism*; D008854:Microscopy, Electron / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D064448:Filamins; D008840:Microfilament Proteins