Microtubules as platforms for assaying actin polymerization in vivo.

The actin cytoskeleton is continuously remodeled through cycles of actin filament assembly and disassembly. Filaments are born through nucleation and shaped into supramolecular structures with various essential functions. These range from contractile and protrusive assemblies in muscle and non-muscl...

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Main Authors: J Margit Oelkers, Marlene Vinzenz, Maria Nemethova, Sonja Jacob, Frank P L Lai, Jennifer Block, Malgorzata Szczodrak, Eugen Kerkhoff, Steffen Backert, Kai Schlüter, Theresia E B Stradal, J Victor Small, Stefan A Koestler, Klemens Rottner
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0019931&type=printable
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author J Margit Oelkers
Marlene Vinzenz
Maria Nemethova
Sonja Jacob
Frank P L Lai
Jennifer Block
Malgorzata Szczodrak
Eugen Kerkhoff
Steffen Backert
Kai Schlüter
Theresia E B Stradal
J Victor Small
Stefan A Koestler
Klemens Rottner
author_facet J Margit Oelkers
Marlene Vinzenz
Maria Nemethova
Sonja Jacob
Frank P L Lai
Jennifer Block
Malgorzata Szczodrak
Eugen Kerkhoff
Steffen Backert
Kai Schlüter
Theresia E B Stradal
J Victor Small
Stefan A Koestler
Klemens Rottner
author_sort J Margit Oelkers
collection DOAJ
description The actin cytoskeleton is continuously remodeled through cycles of actin filament assembly and disassembly. Filaments are born through nucleation and shaped into supramolecular structures with various essential functions. These range from contractile and protrusive assemblies in muscle and non-muscle cells to actin filament comets propelling vesicles or pathogens through the cytosol. Although nucleation has been extensively studied using purified proteins in vitro, dissection of the process in cells is complicated by the abundance and molecular complexity of actin filament arrays. We here describe the ectopic nucleation of actin filaments on the surface of microtubules, free of endogenous actin and interfering membrane or lipid. All major mechanisms of actin filament nucleation were recapitulated, including filament assembly induced by Arp2/3 complex, formin and Spir. This novel approach allows systematic dissection of actin nucleation in the cytosol of live cells, its genetic re-engineering as well as screening for new modifiers of the process.
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publisher Public Library of Science (PLoS)
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series PLoS ONE
spelling doaj-art-d34b14bc521442dea6a348502f3dc5432025-08-20T02:08:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0165e1993110.1371/journal.pone.0019931Microtubules as platforms for assaying actin polymerization in vivo.J Margit OelkersMarlene VinzenzMaria NemethovaSonja JacobFrank P L LaiJennifer BlockMalgorzata SzczodrakEugen KerkhoffSteffen BackertKai SchlüterTheresia E B StradalJ Victor SmallStefan A KoestlerKlemens RottnerThe actin cytoskeleton is continuously remodeled through cycles of actin filament assembly and disassembly. Filaments are born through nucleation and shaped into supramolecular structures with various essential functions. These range from contractile and protrusive assemblies in muscle and non-muscle cells to actin filament comets propelling vesicles or pathogens through the cytosol. Although nucleation has been extensively studied using purified proteins in vitro, dissection of the process in cells is complicated by the abundance and molecular complexity of actin filament arrays. We here describe the ectopic nucleation of actin filaments on the surface of microtubules, free of endogenous actin and interfering membrane or lipid. All major mechanisms of actin filament nucleation were recapitulated, including filament assembly induced by Arp2/3 complex, formin and Spir. This novel approach allows systematic dissection of actin nucleation in the cytosol of live cells, its genetic re-engineering as well as screening for new modifiers of the process.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0019931&type=printable
spellingShingle J Margit Oelkers
Marlene Vinzenz
Maria Nemethova
Sonja Jacob
Frank P L Lai
Jennifer Block
Malgorzata Szczodrak
Eugen Kerkhoff
Steffen Backert
Kai Schlüter
Theresia E B Stradal
J Victor Small
Stefan A Koestler
Klemens Rottner
Microtubules as platforms for assaying actin polymerization in vivo.
PLoS ONE
title Microtubules as platforms for assaying actin polymerization in vivo.
title_full Microtubules as platforms for assaying actin polymerization in vivo.
title_fullStr Microtubules as platforms for assaying actin polymerization in vivo.
title_full_unstemmed Microtubules as platforms for assaying actin polymerization in vivo.
title_short Microtubules as platforms for assaying actin polymerization in vivo.
title_sort microtubules as platforms for assaying actin polymerization in vivo
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0019931&type=printable
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