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The bacterial virulence factors VopL and VopF nucleate actin from the pointed end.


ABSTRACT: VopL and VopF (VopL/F) are tandem WH2-domain actin assembly factors used by infectious Vibrio species to induce actin assembly in host cells. There is disagreement about the filament assembly mechanism of VopL/F, including whether they associate with the filament barbed or pointed end. Here, we used multicolor total internal reflection fluorescence microscopy to directly observe actin assembly with fluorescently labeled VopL/F. In actin monomer assembly reactions, VopL/F exclusively nucleate actin filament assemblies, remaining only briefly associated with the pointed end. VopL/F do not associate with the ends of preassembled filaments. In assembly reactions with saturating profilin, ?85% of VopL/F molecules also promote nucleation from the pointed end, whereas a smaller fraction (<15%) associate for ?25 s with the barbed end of preassembled filaments, inhibiting their elongation. We conclude that VopL/F function primarily as actin nucleation factors that remain briefly (?100 s) associated with the pointed end.

SUBMITTER: Burke TA 

PROVIDER: S-EPMC5412564 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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The bacterial virulence factors VopL and VopF nucleate actin from the pointed end.

Burke Thomas A TA   Harker Alyssa J AJ   Dominguez Roberto R   Kovar David R DR  

The Journal of cell biology 20170331 5


VopL and VopF (VopL/F) are tandem WH2-domain actin assembly factors used by infectious <i>Vibrio</i> species to induce actin assembly in host cells. There is disagreement about the filament assembly mechanism of VopL/F, including whether they associate with the filament barbed or pointed end. Here, we used multicolor total internal reflection fluorescence microscopy to directly observe actin assembly with fluorescently labeled VopL/F. In actin monomer assembly reactions, VopL/F exclusively nucle  ...[more]

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