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Microtubules as platforms for assaying actin polymerization in vivo.
Oelkers, J Margit Vinzenz, Marlene Nemethova, Maria Jacob, Sonja Lai, Frank P L Block, Jennifer Szczodrak, Malgorzata Kerkhoff, Eugen Backert, Steffen Schlüter, Kai
Oelkers, J Margit
Vinzenz, Marlene
Nemethova, Maria
Jacob, Sonja
Lai, Frank P L
Block, Jennifer
Szczodrak, Malgorzata
Kerkhoff, Eugen
Backert, Steffen
Schlüter, Kai
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Abstract
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.
Citation
Microtubules as platforms for assaying actin polymerization in vivo. 2011, 6 (5):e19931 PLoS ONE
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Journal
PubMed ID
PubMed Central ID
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Article
Language
en
Description
Series/Report no.
ISSN
1932-6203