Actin Filament Barbed-End Depolymerization by Combined Action of Profilin, Cofilin, and Twinfilin

Abstract

Cellular actin dynamics results from the collective action of hundreds of regulatory proteins, majority of which target actin filaments at their barbed ends. Three key actin binding proteins—profilin, cofilin, and twinfilin—individually depolymerize filament barbed ends. Notwithstanding recent leaps in our understanding of their individual action, how they collectively regulate filament dynamics remains an open question. In the absence of direct and simultaneous visualization of these proteins at barbed ends, gaining mechanistic insights has been challenging. We have here investigated multicomponent dynamics of profilin, cofilin, and twinfilin using a hybrid approach that combines high-throughput single filament experiments with theory. We discovered that while twinfilin competes with profilin, it promotes binding of cofilin to filament sides. Interestingly, contrary to previous expectations, we found that profilin and cofilin can simultaneously bind the same filament barbed end, resulting in its accelerated depolymerization. Our study reveals that pairwise interactions can effectively capture depolymerization dynamics in simultaneous presence of all three proteins. We thus believe that our approach of employing a theory-experiment dialog can potentially help decipher multicomponent regulation of actin dynamics. Published by the American Physical Society 2024