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

Karen L. Anderson; Christopher Page; Mark F. Swift; Praveen Suraneni; Mandy E.W. Janssen; Thomas D. Pollard; Rong Li; Niels Volkmann; Dorit Hanein

doi:10.1016/j.jsb.2016.12.010

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

Karen L. Anderson, Christopher Page, Mark F. Swift, Praveen Suraneni, Mandy E.W. Janssen, Thomas D. Pollard, Rong Li, Niels Volkmann, Dorit Hanein

School of Medicine

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

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.

Original language	English (US)
Pages (from-to)	312-321
Number of pages	10
Journal	Journal of Structural Biology
Volume	197
Issue number	3
DOIs	https://doi.org/10.1016/j.jsb.2016.12.010
State	Published - Mar 1 2017

Keywords

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

ASJC Scopus subject areas

Structural Biology

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10.1016/j.jsb.2016.12.010

Cite this

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title = "Nano-scale actin-network characterization of fibroblast cells lacking functional Arp2/3 complex",

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",

author = "Anderson, {Karen L.} and Christopher Page and Swift, {Mark F.} and Praveen Suraneni and Janssen, {Mandy E.W.} and Pollard, {Thomas D.} and Rong Li and Niels Volkmann and Dorit Hanein",

note = "Funding Information: This work was supported by NIH program project grants P01 GM066311 (TDP, RL) and P01 GM098412 (DH, NV). NIH grant S10 OD012372 (DH) and P01 GM098412-S1 (DH) funded the purchase of the Titan Krios TEM and Falcon II direct detection imaging device. DH is grateful for time provided on the Titan Krios at the Centre for Bioimaging Science at the National University of Singapore (Dr. P. Matsudaira), and on the Polara (FEI company) of the Cryo-Electron Microscopy Facility at the Broad Center, Caltech (Dr. G. Jensen). Special thanks to Drs. Songye Chen, Caltech and Myint Aung, National University of Singapore. Publisher Copyright: {\textcopyright} 2016 Elsevier Inc.",

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doi = "10.1016/j.jsb.2016.12.010",

language = "English (US)",

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journal = "Journal of Structural Biology",

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AU - Anderson, Karen L.

AU - Page, Christopher

AU - Swift, Mark F.

AU - Suraneni, Praveen

AU - Janssen, Mandy E.W.

AU - Pollard, Thomas D.

AU - Li, Rong

AU - Volkmann, Niels

AU - Hanein, Dorit

N1 - Funding Information: This work was supported by NIH program project grants P01 GM066311 (TDP, RL) and P01 GM098412 (DH, NV). NIH grant S10 OD012372 (DH) and P01 GM098412-S1 (DH) funded the purchase of the Titan Krios TEM and Falcon II direct detection imaging device. DH is grateful for time provided on the Titan Krios at the Centre for Bioimaging Science at the National University of Singapore (Dr. P. Matsudaira), and on the Polara (FEI company) of the Cryo-Electron Microscopy Facility at the Broad Center, Caltech (Dr. G. Jensen). Special thanks to Drs. Songye Chen, Caltech and Myint Aung, National University of Singapore. Publisher Copyright: © 2016 Elsevier Inc.

PY - 2017/3/1

Y1 - 2017/3/1

N2 - 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.

AB - 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.

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KW - Quantitative automated analysis

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

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Keywords

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