Turnover of branched actin filament networks by stochastic fragmentation with ADF/cofilin

Mol Biol Cell. 2011 Jul 15;22(14):2541-50. doi: 10.1091/mbc.E11-01-0052. Epub 2011 May 25.

Abstract

Cell motility depends on the rapid assembly, aging, severing, and disassembly of actin filaments in spatially distinct zones. How a set of actin regulatory proteins that sustains actin-based force generation during motility work together in space and time remains poorly understood. We present our study of the distribution and dynamics of Arp2/3 complex, capping protein (CP), and actin-depolymerizing factor (ADF)/cofilin in actin "comet tails," using a minimal reconstituted system with nucleation-promoting factor (NPF)-coated beads. The Arp2/3 complex concentrates at nucleation sites near the beads as well as in the first actin shell. CP colocalizes with actin and is homogeneously distributed throughout the comet tail; it serves to constrain the spatial distribution of ATP/ADP-P(i) filament zones to areas near the bead. The association of ADF/cofilin with the actin network is therefore governed by kinetics of actin assembly, actin nucleotide state, and CP binding. A kinetic simulation accurately validates these observations. Following its binding to the actin networks, ADF/cofilin is able to break up the dense actin filament array of a comet tail. Stochastic severing by ADF/cofilin loosens the tight entanglement of actin filaments inside the comet tail and facilitates turnover through the macroscopic release of large portions of the aged actin network.

Publication types

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

MeSH terms

  • Actin Capping Proteins / chemistry
  • Actin Cytoskeleton / chemistry*
  • Actin-Related Protein 2-3 Complex / chemistry*
  • Animals
  • Cattle
  • Cell Movement / physiology
  • Destrin / chemistry*
  • Fluorescence Recovery After Photobleaching
  • Kinetics
  • Microspheres
  • Rabbits

Substances

  • Actin Capping Proteins
  • Actin-Related Protein 2-3 Complex
  • Destrin