Phosphorylation of the Arp2 subunit relieves auto-inhibitory interactions for Arp2/3 complex activation

PLoS Comput Biol. 2011 Nov;7(11):e1002226. doi: 10.1371/journal.pcbi.1002226. Epub 2011 Nov 10.


Actin filament assembly by the actin-related protein (Arp) 2/3 complex is necessary to build many cellular structures, including lamellipodia at the leading edge of motile cells and phagocytic cups, and to move endosomes and intracellular pathogens. The crucial role of the Arp2/3 complex in cellular processes requires precise spatiotemporal regulation of its activity. While binding of nucleation-promoting factors (NPFs) has long been considered essential to Arp2/3 complex activity, we recently showed that phosphorylation of the Arp2 subunit is also necessary for Arp2/3 complex activation. Using molecular dynamics simulations and biochemical assays with recombinant Arp2/3 complex, we now show how phosphorylation of Arp2 induces conformational changes permitting activation. The simulations suggest that phosphorylation causes reorientation of Arp2 relative to Arp3 by destabilizing a network of salt-bridge interactions at the interface of the Arp2, Arp3, and ARPC4 subunits. Simulations also suggest a gain-of-function ARPC4 mutant that we show experimentally to have substantial activity in the absence of NPFs. We propose a model in which a network of auto-inhibitory salt-bridge interactions holds the Arp2 subunit in an inactive orientation. These auto-inhibitory interactions are destabilized upon phosphorylation of Arp2, allowing Arp2 to reorient to an activation-competent state.

Publication types

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

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / metabolism
  • Actin-Related Protein 2 / chemistry
  • Actin-Related Protein 2 / genetics
  • Actin-Related Protein 2 / metabolism*
  • Actin-Related Protein 2-3 Complex / chemistry
  • Actin-Related Protein 2-3 Complex / genetics
  • Actin-Related Protein 2-3 Complex / metabolism*
  • Humans
  • Molecular Dynamics Simulation
  • Mutation
  • Phosphorylation
  • Polymerization
  • Protein Conformation
  • Protein Processing, Post-Translational*


  • ACTR2 protein, human
  • Actin-Related Protein 2
  • Actin-Related Protein 2-3 Complex