Cofilin-Mediated Filament Softening and Crosslinking Counterbalance to Enhance Actin Network Flexibility

Filamentous-actin (F-actin) crosslinking within the cell cytoskeleton mediates the transmission of mechanical forces, enabling changes in cell shape, as occurs during cell division and cell migration. Crosslinking by actin binding proteins (ABPs) generally increases the connectivity of the F-actin network, but also increases network rigidity. As a result, there is a narrow range in the concentration of crosslinker protein at which F-actin networks are both connected and labile. Another ABP, cofilin, severs F-actin filaments at high pH through increasing their bending flexibility and concentrating mechanical stress, inducing fragmentation. By contrast, at lower pH, cofilin increases filament flexibility yet does not sever. Instead, it forms disulfide bonds, which crosslink F-actin into bundles, and bundles into networks. Here, we combine light microscopy and rheology to determine the impact of two potentially opposing effects on the mechanics of F-actin networks-increased flexibility at the filament level, and increased connectivity at the network level. Indeed, by linear rheology, we find that these mechanisms are counterbalanced, such that cofilactin network moduli are only weakly dependent on cofilin concentration over a broad range, in contrast to the dramatic stiffening that occurs with F-actin crosslinking protein. Further, by nonlinear rheology, the network stiffens at a higher stress than crosslinking protein, indicative of a broader range in which the material remains flexible. These results may enable F-actin networks to increase connectivity without heavy penalties to rigidity, and thus provide a new route to modulating active polymer mechanics unseen using traditional F-actin accessory proteins.