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

Nature. 2000 Apr 27;404(6781):1007-11. doi: 10.1038/35010008.

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 membrane. 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 forward. 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 proposed 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acanthamoeba
  • Actin Depolymerizing Factors
  • Actin-Related Protein 2
  • Actin-Related Protein 3
  • Actins / metabolism*
  • Animals
  • Cattle
  • Contractile Proteins*
  • Cytoskeletal Proteins*
  • Destrin
  • Humans
  • Microfilament Proteins / metabolism
  • Muscle, Skeletal / metabolism
  • Nucleotides / metabolism
  • Profilins
  • Proteins / metabolism*
  • Rabbits
  • Recombinant Proteins / metabolism
  • Wiskott-Aldrich Syndrome Protein
  • Wiskott-Aldrich Syndrome Protein Family

Substances

  • ACTR2 protein, human
  • ACTR3 protein, human
  • Actin Depolymerizing Factors
  • Actin-Related Protein 2
  • Actin-Related Protein 3
  • Actins
  • Contractile Proteins
  • Cytoskeletal Proteins
  • Destrin
  • Microfilament Proteins
  • Nucleotides
  • PFN1 protein, human
  • Profilins
  • Proteins
  • Recombinant Proteins
  • WAS protein, human
  • WASF1 protein, human
  • Wiskott-Aldrich Syndrome Protein
  • Wiskott-Aldrich Syndrome Protein Family