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ActA and human zyxin harbour Arp2/3-independent actin-polymerization activity

Nature Cell Biology volume 3, pages 699–707 (2001)Cite this article

Abstract

The actin cytoskeleton is a dynamic network that is composed of a variety of F-actin structures. To understand how these structures are produced, we tested the capacity of proteins to direct actin polymerization in a bead assay in vitro and in a mitochondrial-targeting assay in cells. We found that human zyxin and the related protein ActA of Listeria monocytogenes can generate new actin structures in a vasodilator-stimulated phosphoprotein-dependent (VASP) manner, but independently of the Arp2/3 complex. These results are consistent with the concept that there are multiple actin-polymerization machines in cells. With these simple tests it is possible to probe the specific function of proteins or identify novel molecules that act upon cellular actin polymerization.

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Figure 1: Structural features of the ActA, WASP and zyxin proteins.
Figure 2: ActA-Pro-coated beads promote actin polymerization in cell-free extracts.
Figure 3: The Arp2/3 complex is not recruited to the surface of ActA-Pro beads.
Figure 4: ActA-Pro-directed actin polymerization is independent of the Arp2/3 complex.
Figure 5: ActA-Pro reduces actin polymerization at cell-adhesion sites.
Figure 6: The proline-rich domain of zyxin mediates actin polymerization in cells.
Figure 7: Zyxin-mediated actin polymerization is independent of the Arp2/3 complex.
Figure 8: Zyxin-mediated actin polymerization is dependent on VASP.

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Acknowledgements

We thank L. Cabanié for technical support and members of the Louvard laboratory and M. Beckerle for valuable discussions. We also thank P. Chavrier, P. Cossart, E. Gouin, L. Hoffman, C. Koehler and L. Machesky for generously providing reagents. This work was supported by a Curie Fellowship (to R.M.G. and J.P.), a MENRT fellowship (to J.F. and V.N.), and grants from the Association pour la Recherche sur le Cancer and the Centre National de la Recherche Scientifique (France).

Author information

Author notes

  1. Vincent Noireaux

    Present address: The Rockefeller University, Center for Studies in Physics and Biology, Box 265, 1230 York Avenue, New York, NY 10021, USA

  2. Roy M. Golsteyn

    Present address: Division de Cancérologie Expérimentale, Institut de Recherches Servier, 125, chemin de Ronde, 78290, Croissy-sur-Seine, France

  3. Evelyne Friederich

    Present address: CRP-Santé, 18 rue Dicks, L-1417, Luxembourg

  4. Julie Fradelizi and Vincent Noireaux: These authors contributed equally to this work

  5. Roy M. Golsteyn and Evelyne Friederich: These authors share last authorship

Authors and Affiliations

  1. Laboratoire de Morphogenèse et Signalisation Cellulaires, Unité Mixte de Recherche CNRS/Institut Curie (UMR144), 26 rue d'Ulm, Paris, cedex 05, 75248, France

    Julie Fradelizi, Bernadette Menichi, Daniel Louvard, Roy M. Golsteyn & Evelyne Friederich

  2. Laboratoire Physico-Chimie Curie, Unité Mixte de Recherche CNRS/Institut Curie (UMR168), 26 rue d'Ulm, Paris, cedex 05, 75248, France

    Vincent Noireaux, Julie Plastino & Cécile Sykes

  3. Laboratoire Franco-Luxembourgeois de Recherche Biomédicale, CNRS/CRP-Santé, Centre Universitaire, 162A, av. de la Faïencerie, L-1511, Luxembourg

    Evelyne Friederich

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Supplementary information

Supplementary figure

Figure S1 Effect of cytochalasin D on the ATPase activity of actin during polymerization. (PDF 26 kb)

Figure S2 Inhibition of actin polymerization on the surface of WA or ActA-Pro-beads by increasing concentrations of latrunculin.

Figure S3 Quantification of fluorescence associated with WA or ActA-Pro beads immunostained for VASP or Arp3 in the absence of latrunculin A.

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Fradelizi, J., Noireaux, V., Plastino, J. et al. ActA and human zyxin harbour Arp2/3-independent actin-polymerization activity. Nat Cell Biol 3, 699–707 (2001). https://doi.org/10.1038/35087009

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