Motor-driven modulation of actin network mechanics across linear and nonlinear regimes

Cytoskeletal networks enable cells to dynamically regulate their mechanical properties in response to internal forces and external cues. Here, we investigate how motor activity influences the structure and mechanics of actomyosin networks reconstituted in vitro from filamentous actin, myosin II minifilaments, and transient -actinin cross-linkers. By varying the myosin-to-actin molar ratio (RMA), we observe a transition from isotropic actin meshes to contractile, coarsened architectures marked by bundled filaments and increasing spatial correlation lengths ({xi}z,{xi} t). Optical tweezers microrheology reveals a nonmonotonic mechanical response: at low RMA, networks fluidize, with reductions in the plateau modulus (G0), zero-shear viscosity (0), and fast relaxation timescales ([Formula],{tau} 1). At higher motor levels, the networks stiffen and retain internal stress, reflecting contractile reinforcement. Notably,{tau} 1 exhibits a minimum when plotted against{xi} z, suggesting that intermediate levels of coarsening facilitate efficient local stress dissipation. These results identify distinct mechanical regimes governed by motor-induced remodeling and highlight a structural basis for the dual roles of myosin in fluidization and reinforcement.