The Membrane-Associated Protein, Supervillin, Accelerates F-Actin-Dependent Rapid Integrin Recycling and Cell Motility

Authors

  • Zhiyou Fang,

    1. Department of Cell Biology, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    Search for more papers by this author
  • Norio Takizawa,

    1. Department of Cell Biology, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    Search for more papers by this author
  • Korey A. Wilson,

    1. National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA
    Search for more papers by this author
  • Tara C. Smith,

    1. Department of Cell Biology, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    Search for more papers by this author
  • Anna Delprato,

    1. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Program in Molecular Medicine, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    3. Current address: INSERM U869, Institut de Chimie et Biologie, 2 rue, Robert Escarpit, F-33607 Pessac, France
    Search for more papers by this author
  • Michael W. Davidson,

    1. National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA
    Search for more papers by this author
  • David G. Lambright,

    1. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Program in Molecular Medicine, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    Search for more papers by this author
  • Elizabeth J. Luna

    Corresponding author
    1. Department of Cell Biology, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    2. Cell and Developmental Dynamics Program, University of Massachusetts Medical School, Biotech 4, Suite 306, 377 Plantation Street, Worcester, MA 01605, USA
    Search for more papers by this author

Elizabeth J. Luna, Elizabeth.Luna@umassmed.edu

Abstract

In migrating cells, the cytoskeleton coordinates signal transduction and redistribution of transmembrane proteins, including integrins and growth factor receptors. Supervillin is an F-actin- and myosin II-binding protein that tightly associates with signaling proteins in cholesterol-rich, ‘lipid raft’ membrane microdomains. We show here that supervillin also can localize with markers for early and sorting endosomes (EE/SE) and with overexpressed components of the Arf6 recycling pathway in the cell periphery. Supervillin tagged with the photoswitchable fluorescent protein, tdEos, moves both into and away from dynamic structures resembling podosomes at the basal cell surface. Rapid integrin recycling from EE/SE is inhibited in supervillin-knockdown cells, but the rates of integrin endocytosis and recycling from the perinuclear recycling center (PNRC) are unchanged. A lack of synergy between supervillin knockdown and the actin filament barbed-end inhibitor, cytochalasin D, suggests that both treatments affect actin-dependent rapid recycling. Supervillin also enhances signaling from the epidermal growth factor receptor (EGFR) to extracellular signal-regulated kinases (ERKs) 1 and 2 and increases the velocity of cell translocation. These results suggest that supervillin, F-actin and associated proteins coordinate a rapid, basolateral membrane recycling pathway that contributes to ERK signaling and actin-based cell motility.

Ancillary