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Significance Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) proteins are required for the formation and maintenance of filopodia, finger-like projections at the leading edge of migrating cells that are composed of parallel actin filaments bundled by Fascin. We imaged individual fluorescently labeledDrosophila Ena molecules on both single and Fascin-bundled actin filaments in vitro. Ena stimulates actin assembly by remaining continuously associated with the barbed end and increasing the elongation rate by approximately two- to threefold. Remarkably, the frequency and length of Ena’s processive runs are enhanced on filaments within a Fascin bundle, which drives a positive feedback cycle that allows the assembly of uniformly thick filopodia-like F-actin bundles composed of multiple filaments with aligned ends.

Ena/VASP Enabled is a highly processive actin polymerase tailored to self-assemble parallel-bundled F-actin networks with Fascin