Processive acceleration of actin barbed-end assembly by N-WASP

Supplemental Materials

This article contains the following supporting material:

  • Supplemental Materials
  • Movie01 - Supplementary Movie 1. Single filament capture is non-processive. Conditions as in Figure 1. Time-lapse TIRF microcopy movie of elongating barbed ends (arrowhead) attached to a GSTPP- WWCA coated nanofiber. Barbed ends polymerized either parallel to the nanofiber long axis at the diffusion-limited rate (white arrowhead) or perpendicular to the nanofiber at a substantially reduced rate (black arrowhead).
  • Movie02 - Supplementary Movie 2. Rapid processive elongation in standard magnesium. Conditions as in Figure 2. Time-lapse TIRF microscopy movie of sustained processive elongation of actin filaments on GST-PP-WWCA coated glass nanofibers (black arrowhead) in 1 mM Mg2+ with barbed ends attached to nanofibers (white arrowhead).
  • Movie03 - Supplementary Movie 3. Rapid processive elongation in high magnesium. Conditions as in Figure 2. Time-lapse TIRF microscopy movie of sustained rapid elongation of actin filaments (black arrowhead) with barbed ends attached to GST-PP-WWCA nanofibers in 10 mM Mg2+.
  • Movie04 - Supplementary Movie 4. Rapid processive elongation in standard magnesium. Conditions as in Figure 2 except with 10 mM total Mg2+. Time-lapse TIRF microscopy movie of two elongating actin filaments (black arrowheads). Filament barbed ends are bundled (open arrowhead) and lead to a nearby GST-PP-WWCA nanofibers. The lower filament undergoes sustained, rapid elongation, while the upper filament elongates at the normal rate.
  • Movie05 - Supplementary Movie 5. Profilin slows rate of bundle mediated processive elongation. Conditions as in Figure 3. Time-lapse TIRF microcopy movie of sustained processive association of profilin-actin with actin filaments barbed ends attached to GST-PP-WWCA coated nanofibers resulting in filament buckling (black arrowhead).
  • Movie06 - Supplementary Movie 6. Filaments do not interact with BSA blocked nanofibers. Conditions as in Supplementary Figure 1. Time-lapse TIRF microcopy movie of barbed end of actin filaments (black arrowhead) growing past FITC-BSA coated nanofibers.