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

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

Author information

1
Bioinformatics and Structural Biology Program, Sanford-Burnham Medical Research Institute, La Jolla, CA, United States.
2
Department of Cell Biology, Johns Hopkins School of Medicine, Baltimore, MD, United States.
3
Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, United States; Department of Cell Biology and of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, United States.
4
Bioinformatics and Structural Biology Program, Sanford-Burnham Medical Research Institute, La Jolla, CA, United States. Electronic address: niels@burnham.org.
5
Bioinformatics and Structural Biology Program, Sanford-Burnham Medical Research Institute, La Jolla, CA, United States. Electronic address: dorit@burnham.org.

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

PMID:
28013022
PMCID:
PMC5360490
DOI:
10.1016/j.jsb.2016.12.010
[Indexed for MEDLINE]
Free PMC Article

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