Abstract
The actin cytoskeleton is a dynamic network that is composed of a variety of F-actin structures. To understand how these structures are produced, we tested the capacity of proteins to direct actin polymerization in a bead assay in vitro and in a mitochondrial-targeting assay in cells. We found that human zyxin and the related protein ActA of Listeria monocytogenes can generate new actin structures in a vasodilator-stimulated phosphoprotein-dependent (VASP) manner, but independently of the Arp2/3 complex. These results are consistent with the concept that there are multiple actin-polymerization machines in cells. With these simple tests it is possible to probe the specific function of proteins or identify novel molecules that act upon cellular actin polymerization.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Actin Cytoskeleton / drug effects
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Actin Cytoskeleton / metabolism*
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Actin Cytoskeleton / ultrastructure
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Actin-Related Protein 2
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Actin-Related Protein 3
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Actins / metabolism*
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Animals
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Bacterial Proteins / metabolism*
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Biological Assay
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Cell Adhesion Molecules / metabolism
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Cell-Free System
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Chlorocebus aethiops
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Cytoskeletal Proteins*
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Fluorescent Antibody Technique
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Glycoproteins
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HeLa Cells / cytology
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HeLa Cells / drug effects
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HeLa Cells / metabolism
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Humans
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Membrane Proteins / metabolism*
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Metalloproteins / genetics
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Metalloproteins / metabolism*
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Microfilament Proteins
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Microspheres
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Mitochondria / metabolism
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Mitochondria / ultrastructure
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Phosphoproteins / metabolism
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Polymers / metabolism*
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Proteins / metabolism
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Recombinant Proteins / metabolism
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Transfection
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Vasodilator-Stimulated Phosphoprotein
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Vero Cells / cytology
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Vero Cells / drug effects
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Vero Cells / metabolism
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Wiskott-Aldrich Syndrome Protein
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Zyxin
Substances
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ACTR2 protein, human
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ACTR3 protein, human
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Actin-Related Protein 2
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Actin-Related Protein 3
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Actins
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Bacterial Proteins
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Cell Adhesion Molecules
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Cytoskeletal Proteins
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Glycoproteins
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Membrane Proteins
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Metalloproteins
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Microfilament Proteins
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Phosphoproteins
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Polymers
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Proteins
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Recombinant Proteins
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WAS protein, human
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Wiskott-Aldrich Syndrome Protein
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ZYX protein, human
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Zyxin
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Vasodilator-Stimulated Phosphoprotein
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actA protein, Listeria monocytogenes