Nano-scale actin-network characterization of fibroblast cells lacking functional Arp2/3 complex

J Struct Biol. 2017 Mar;197(3):312-321. doi: 10.1016/j.jsb.2016.12.010. Epub 2016 Dec 21.

Abstract

Arp2/3 complex is thought to be the primary protrusive force generator in cell migration by controlling the assembly and turnover of the branched filament network that pushes the leading edge of moving cells forward. However, mouse fibroblasts without functional Arp2/3 complex migrate at rates similar to wild-type cells, contradicting this paradigm. We show by correlative fluorescence and large-scale cryo-tomography studies combined with automated actin-network analysis that the absence of functional Arp2/3 complex has profound effects on the nano-scale architecture of actin networks. Our quantitative analysis at the single-filament level revealed that cells lacking functional Arp2/3 complex fail to regulate location-dependent fine-tuning of actin filament growth and organization that is distinct from its role in the formation and regulation of dendritic actin networks.

Keywords: Actin networks; Correlative imaging; Large-scale cellular cryo-tomography; Quantitative automated analysis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actin-Related Protein 2-3 Complex / genetics
  • Actin-Related Protein 2-3 Complex / metabolism*
  • Actins / metabolism*
  • Actins / ultrastructure
  • Animals
  • Cells, Cultured
  • Cryoelectron Microscopy
  • Electron Microscope Tomography
  • Fibroblasts / metabolism*
  • Fibroblasts / ultrastructure
  • Mice

Substances

  • Actin-Related Protein 2-3 Complex
  • Actins