Reconstitution of the transition from lamellipodium to filopodium in a membrane-free system

Lior Haviv, Yifat Brill-Karniely, Rachel Mahaffy, Frederic Backouche, Avinoam Ben-Shaul, Thomas D. Pollard, Anne Bernheim-Groswasser

Research output: Contribution to journalArticlepeer-review

63 Scopus citations

Abstract

The cellular cytoskeleton is a complex dynamical network that constantly remodels as cells divide and move. This reorganization process occurs not only at the cell membrane, but also in the cell interior (bulk). During locomotion, regulated actin assembly near the plasma membrane produces lamellipodia and filopodia. Therefore, most in vitro experiments explore phenomena taking place in the vicinity of a surface. To understand how the molecular machinery of a cell self-organizes in a more general way, we studied bulk polymerization of actin in the presence of actin-related protein 2/3 complex and a nucleation promoting factor as a model for actin assembly in the cell inferior separate from membranes. Bulk polymerization of actin in the presence of the verprolin homology, cofilin homology, and acidic region, domain of Wiskott-Aldrich syndrome protein, and actin-related protein 2/3 complex results in spontaneous formation of diffuse aster-like structures. In the presence of fascin these asters transition into stars with bundles of actin filaments growing from the surface, similar to star-like structures recently observed in vivo. The transition from asters to stars depends on the ratio [fascin]/[G actin]. The polarity of the actin filaments during the transition is preserved, as in the transition from lamellipodia to filopodia. Capping protein inhibits star formation. Based on these experiments and kinetic Monte Carlo simulations, we propose a model for the spontaneous self-assembly of asters and their transition into stars. This mechanism may apply to the transition from lamellipodia to filopodia in vivo.

Original languageEnglish
Pages (from-to)4906-4911
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number13
DOIs
StatePublished - 28 Mar 2006

Keywords

  • Actin self-assembly
  • Asters
  • Cellular protrusions
  • Monte Carlo simulations
  • Stars

ASJC Scopus subject areas

  • General

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