4.4 Article

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

Journal

JOURNAL OF STRUCTURAL BIOLOGY
Volume 197, Issue 3, Pages 312-321

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2016.12.010

Keywords

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

Funding

  1. NIH [P01 GM066311, P01 GM098412, S10 OD012372, P01 GM098412-S1]
  2. DH, NV [S10 OD012372]

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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 cryotomography 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. (C) 2016 Elsevier Inc. All rights reserved.

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