Reconstitution of actin-based motility of Listeria and Shigella using pure proteins

Nature. 1999 Oct 7;401(6753):613-6. doi: 10.1038/44183.

Abstract

Actin polymerization is essential for cell locomotion and is thought to generate the force responsible for cellular protrusions. The Arp2/3 complex is required to stimulate actin assembly at the leading edge in response to signalling. The bacteria Listeria and Shigella bypass the signalling pathway and harness the Arp2/3 complex to induce actin assembly and to propel themselves in living cells. However, the Arp2/3 complex alone is insufficient to promote movement. Here we have used pure components of the actin cytoskeleton to reconstitute sustained movement in Listeria and Shigella in vitro. Actin-based propulsion is driven by the free energy released by ATP hydrolysis linked to actin polymerization, and does not require myosin. In addition to actin and activated Arp2/3 complex, actin depolymerizing factor (ADF, or cofilin) and capping protein are also required for motility as they maintain a high steady-state level of G-actin, which controls the rate of unidirectional growth of actin filaments at the surface of the bacterium. The movement is more effective when profilin, alpha-actinin and VASP (for Listeria) are also included. These results have implications for our understanding of the mechanism of actin-based motility in cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Depolymerizing Factors
  • Actin-Related Protein 2
  • Actin-Related Protein 3
  • Actins / physiology*
  • Adenosine Triphosphate / physiology
  • Animals
  • Bacterial Proteins
  • Cattle
  • Cell Adhesion Molecules / physiology
  • Contractile Proteins*
  • Cytoskeletal Proteins*
  • DNA-Binding Proteins / physiology
  • Escherichia coli / physiology
  • Humans
  • Listeria monocytogenes / physiology*
  • Microfilament Proteins / physiology
  • Movement
  • Nerve Tissue Proteins / physiology
  • Phosphoproteins / physiology
  • Profilins
  • Rabbits
  • Recombinant Proteins
  • Shigella / physiology*
  • Transcription Factors / physiology
  • Wiskott-Aldrich Syndrome Protein, Neuronal

Substances

  • ACTR2 protein, human
  • ACTR3 protein, human
  • Actin Depolymerizing Factors
  • Actin-Related Protein 2
  • Actin-Related Protein 3
  • Actins
  • Bacterial Proteins
  • Cell Adhesion Molecules
  • Contractile Proteins
  • Cytoskeletal Proteins
  • DNA-Binding Proteins
  • Microfilament Proteins
  • Nerve Tissue Proteins
  • Phosphoproteins
  • Profilins
  • Recombinant Proteins
  • Transcription Factors
  • WASL protein, human
  • Wiskott-Aldrich Syndrome Protein, Neuronal
  • vasodilator-stimulated phosphoprotein
  • virG protein, Shigella flexneri
  • Adenosine Triphosphate