Fig. 1
S. cerevisiae Srv2/CAP and Cofilin synergize to accelerate actin filament depolymerization. a Schematic representation of the experimental strategy. Preformed Alexa-488-labeled actin filaments were captured by coverslip-anchored biotinylated SNAP-CapZ. After 15 min, 1 µM Cof1 and/or 0.5 µM Srv2 (or control buffer) was introduced into the chamber, and depolymerization was monitored. BE barbed end, PE pointed end. b Representative field of view showing anchored filaments aligned under flow, and the methodology used for determining the rate of pointed-end depolymerization from the slope of kymographs. Scale bar, 10 µm. c Rates (±sd) of pointed-end depolymerization in the presence of 1 μM Cof1 and/or 0.5 μM Srv2. Right: Magnified view of Control, Cof1, and Srv2 data. *statistical comparison by two-sample t test against Control (p < 0.05). Number of filament ends analyzed for each condition (left to right): 149, 55, 110 and 37. d Merged two-color kymograph of an Alexa-488-labeled actin filament (green), with 1 μM Cy3-Cof1 (red) introduced at the beginning of the red bar (see Supplementary Movie 1). e Same as (d) but with 1 μM Cy3-Cof1 (red) and 0.5 μM Srv2 (unlabeled). f Rates (±sd) of pointed-end depolymerization by 1 μM Cof1 and/or 0.5 μM Srv2 in the presence of 3 μM G-actin (with or without 6 μM Profilin). *statistical comparison by two-sample t test against Cof1 + Srv2 (p < 0.05). ns no evidence for significance at p = 0.05. Number of filament ends analyzed for each condition (left to right): 64, 83 and 84. Source data are provided as a Source Data file. All experiments were performed at least three independent times, and yielded similar results. Data shown are from one experiment.
