Analysis of the local organization and dynamics of cellular actin networks

J Cell Biol. 2013 Sep 30;202(7):1057-73. doi: 10.1083/jcb.201210123.

Abstract

Actin filaments, with the aid of multiple accessory proteins, self-assemble into a variety of network patterns. We studied the organization and dynamics of the actin network in nonadhesive regions of cells bridging fibronectin-coated adhesive strips. The network was formed by actin nodes associated with and linked by myosin II and containing the formin disheveled-associated activator of morphogenesis 1 (DAAM1) and the cross-linker filamin A (FlnA). After Latrunculin A (LatA) addition, actin nodes appeared to be more prominent and demonstrated drift-diffusion motion. Superresolution microscopy revealed that, in untreated cells, DAAM1 formed patches with a similar spatial arrangement to the actin nodes. Node movement (diffusion coefficient and velocity) in LatA-treated cells was dependent on the level and activity of myosin IIA, DAAM1, and FlnA. Based on our results, we developed a computational model of the dynamic formin-filamin-actin asters that can self-organize into a contractile actomyosin network. We suggest that such networks are critical for connecting distant parts of the cell to maintain the mechanical coherence of the cytoplasm.

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 / physiology*
  • Actomyosin / metabolism
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Blotting, Western
  • Cells, Cultured
  • Computer Simulation
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism*
  • Fetal Proteins / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Filamins / physiology*
  • Fluorescent Antibody Technique
  • Formins
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Immunoenzyme Techniques
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Knockout
  • Microfilament Proteins / antagonists & inhibitors
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Models, Theoretical
  • Myosin Type II / metabolism*
  • Nuclear Proteins / metabolism
  • Phosphoproteins / metabolism
  • RNA, Small Interfering / genetics
  • rho GTP-Binding Proteins / antagonists & inhibitors
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Fetal Proteins
  • Filamins
  • FlnA protein, mouse
  • Formins
  • Lat protein, mouse
  • Membrane Proteins
  • Microfilament Proteins
  • Nuclear Proteins
  • Phosphoproteins
  • RNA, Small Interfering
  • Actomyosin
  • Daam1 protein, mouse
  • Myosin Type II
  • rho GTP-Binding Proteins