Abstract
Rearrangement of actin filaments by polymerization, depolymerization, and severing is important for cell locomotion, membrane trafficking, and many other cellular functions. Cofilin and actin-interacting protein 1 (AIP1; also known as WDR1) are evolutionally conserved proteins that cooperatively sever actin filaments. However, little is known about the biophysical basis of the actin filament severing by these proteins. Here, we performed single-molecule kinetic analyses of fluorescently labeled AIP1 during the severing process of cofilin-decorated actin filaments. Results demonstrated that binding of a single AIP molecule was sufficient to enhance filament severing. After AIP1 binding to a filament, severing occurred with a delay of 0.7 s. Kinetics of binding and dissociation of a single AIP1 molecule to/from actin filaments followed a second-order and a first-order kinetics scheme, respectively. AIP1 binding and severing were detected preferentially at the boundary between the cofilin-decorated and bare regions on actin filaments. Based on the kinetic parameters explored in this study, we propose a possible mechanism behind the enhanced severing by AIP1.