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Direct observation of dendritic actin filament networks nucleated by Arp2/3 complex and WASP/Scar proteins

Author

Listed:
  • Laurent Blanchoin

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

  • Kurt J. Amann

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

  • Henry N. Higgs

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

  • Jean-Baptiste Marchand

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

  • Donald A. Kaiser

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

  • Thomas D. Pollard

    (Structural Biology Laboratory, Salk Institute for Biological Studies)

Abstract

Most nucleated cells crawl about by extending a pseudopod that is driven by the polymerization of actin filaments in the cytoplasm behind the leading edge of the plasma membrane1,2. These actin filaments are linked into a network by Y-branches, with the pointed end of each filament attached to the side of another filament and the rapidly growing barbed end facing forward3. Because Arp2/3 complex nucleates actin polymerization and links the pointed end to the side of another filament in vitro, a dendritic nucleation model has been proposed4 in which Arp2/3 complex initiates filaments from the sides of older filaments. Here we report, by using a light microscopy assay, many new features of the mechanism. Branching occurs during, rather than after, nucleation by Arp2/3 complex activated by the Wiskott–Aldrich syndrome protein (WASP) or Scar protein; capping protein and profilin act synergistically with Arp2/3 complex to favour branched nucleation; phosphate release from aged actin filaments favours dissociation of Arp2/3 complex from the pointed ends of filaments; and branches created by Arp2/3 complex are relatively rigid. These properties result in the automatic assembly of the branched actin network after activation by proteins of the WASP/Scar family and favour the selective disassembly of proximal regions of the network.

Suggested Citation

  • Laurent Blanchoin & Kurt J. Amann & Henry N. Higgs & Jean-Baptiste Marchand & Donald A. Kaiser & Thomas D. Pollard, 2000. "Direct observation of dendritic actin filament networks nucleated by Arp2/3 complex and WASP/Scar proteins," Nature, Nature, vol. 404(6781), pages 1007-1011, April.
    • Handle: RePEc:nat:nature:v:404:y:2000:i:6781:d:10.1038_35010008
    DOI: 10.1038/35010008
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