Cofilin-induced unidirectional cooperative conformational changes in actin filaments revealed by high-speed atomic force microscopy.

Kien Xuan Ngo,Noriyuki Kodera,Taro Qp Uyeda,Toshio Ando,Eisaku Katayama

doi:10.7554/elife.04806

Abstract

High-speed atomic force microscopy was employed to observe structural changes in actin filaments induced by cofilin binding. Consistent with previous electron and fluorescence microscopic studies, cofilin formed clusters along actin filaments, where the filaments were 2-nm thicker and the helical pitch was ~25% shorter, compared to control filaments. Interestingly, the shortened helical pitch was propagated to the neighboring bare zone on the pointed-end side of the cluster, while the pitch on the barbed-end side was similar to the control. Thus, cofilin clusters induce distinctively asymmetric conformational changes in filaments. Consistent with the idea that cofilin favors actin structures with a shorter helical pitch, cofilin clusters grew unidirectionally toward the pointed-end of the filament. Severing was often observed near the boundaries between bare zones and clusters, but not necessarily at the boundaries.

Highlights

Actin filaments are involved in a variety of important functions in eukaryotic cells, including muscle contraction, amoeboid movement, cytokinesis, intracellular transport, and transcriptional regulation within the nucleus
For Atomic force microscope (AFM), actin filaments must be immobilized on the stage, yet they must have the freedom of movement to bind cofilin and exhibit the resultant changes in the helical twist that accompany cofilin binding
A solution of actin filaments was placed on the supported lipid bilayer and high-speed AFM (HS-AFM) was performed

Summary

Introduction

Actin filaments are involved in a variety of important functions in eukaryotic cells, including muscle contraction, amoeboid movement, cytokinesis, intracellular transport, and transcriptional regulation within the nucleus. Three independent biochemical mechanisms are known to inhibit cofilin activity; phosphorylation of Ser, sequestration to phosphatidylinositol 4,5-bisphosphate (PIP2) in plasma membrane, and lower pH (Bernstein and Bamburg, 2010; Bravo-Cordero et al, 2013) These inhibitory mechanisms are implicated in the regulation of cofilin activity during cell migration, since experimental unleashing of inactive cofilin has been shown to initiate cofilin-dependent processes (Bernstein and Bamburg, 2010; Bravo-Cordero et al, 2013).

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