Formins accelerate actin polymerization, assumed to occur through flexible FH1 domain mediated transfer of profilin-actin to the barbed end. To study FH1 properties and address sequence effects including varying length/distribution of profilin-binding proline-rich motifs, we performed all-atom simulations of mouse mDia1, mDia2; budding yeast Bni1, Bnr1; fission yeast Cdc12, For3, and Fus1 FH1s. We find FH1 has flexible regions between high propensity polyproline helix regions. A coarse-grained model retaining sequence-specificity, assuming rigid polyproline segments, describes their size. Multiple bound profilins or profilin-actin complexes expand mDia1-FH1, which may be important in cells. Simulations of the barbed end bound to Bni1-FH1-FH2 dimer show the leading FH1 can better transfer profilin or profilin-actin, having decreasing probability with increasing distance from FH2.

We studied actin filament polymerization and nucleation with molecular dynamics simulations and a previously established coarse-grained model having each residue represented by a single interaction site located at the C α atom. We approximate each actin protein as a fully or partially rigid unit to identify the equilibrium structural ensemble of interprotein complexes. Monomers in the F-actin configuration bound to both barbed and pointed ends of a short F-actin filament at the anticipated locations for polymerization. Binding at both ends occurred with similar affinity. Contacts between residues of the incoming subunit and the short filament were consistent with expectation from models based on crystallography, X-ray diffraction and cryo-electron microscopy. Binding at the barbed and pointed end also occurred at an angle with respect to the polymerizable bound structure, and the angle range depended on the flexibility of the D-loop. Additional barbed end bound states were seen when the incoming subunit was in the G-actin form. Consistent with an activation barrier for pointed end polymerization, G-actin did not bind at an F-actin pointed end. In all cases, binding at the barbed end also occurred in a configuration similar to the antiparallel (lower) dimer. Individual monomers bound each other in a short-pitch helix complex in addition to other configurations, with several of them apparently non-productive for polymerization. Simulations with multiple monomers in the F-actin form show assembly into filaments as well as transient aggregates at the barbed end. We discuss the implications of these observations on the kinetic pathway of actin filament nucleation and polymerization and possibilities for future improvements of the coarse-grained model. SIGNIFICANCE Control of actin filament nucleation and elongation has crucial importance to cellular life. We show that coarse-grained molecular dynamics simulations are a powerful tool which can gauge involved mechanisms at reasonable computational cost, while retaining essential features of the fully atomic, yet less computationally tractable, system. Using a knowledge-based potential demonstrates the power of these methods for explaining and reproducing polymerization. Intermediate actin complexes identified in the simulations may play critical roles in the kinetic pathways of actin polymerization which may have been difficult to observe in prior experiments. These methods have been sparsely applied to the actin system, yet have potential to answer many important questions in the field.

We studied actin filament polymerization and nucleation with molecular dynamics simulations and a previously established coarse-grained model having each residue represented by a single interaction site located at the C α atom. We approximate each actin protein as a fully or partially rigid unit to identify the equilibrium structural ensemble of interprotein complexes. Monomers in the F-actin configuration bound to both barbed and pointed ends of a short F-actin filament at the anticipated locations for polymerization. Binding at both ends occurred with similar affinity. Contacts between residues of the incoming subunit and the short filament were consistent with expectation from models based on crystallography, X-ray diffraction and cryo-electron microscopy. Binding at the barbed and pointed end also occurred at tilted conformations and the barbed end tilt range was dependent on the flexibility of the D-loop. Additional barbed end bound states were seen when the incoming subunit was in the G form. Consistent with an activation barrier for pointed end polymerization, G-actin did not bind at an F-actin pointed end. In all cases, binding at the barbed end also occurred in a configuration similar to the antiparallel (lower) dimer. Individual monomers bound each other in a short-pitch helix complex in addition to other configurations, with several of them apparently non-productive for polymerization. Simulations with multiple monomers in the F-actin form reproduce filamentation as well as transient aggregates at the barbed end. We discuss the implications on the kinetic pathway of actin filament nucleation and polymerization and possibilities for future improvements of the coarse-grained model. SIGNIFICANCE Control of actin filament polymerization and nucleation has crucial importance to cellular life. We show that coarse-grained molecular dynamics simulations are a powerful tool which can gauge involved mechanisms at reasonable computational cost, while retaining essential features of the fully atomic, yet less computationally tractable, system. Using a knowledge-based potential demonstrates the power of these methods at explaining and reproducing polymerization. Intermediate actin complexes identified in the simulations may play critical roles in the kinetic pathways of actin polymerization which may have been difficult to observe in prior experiments. These methods have been sparsely applied to the actin system, yet have potential to answer many important questions in the field.

Formins polymerize actin filaments for the cytokinetic contractile ring. Using in vitro reconstitution of fission yeast contractile ring precursor nodes containing formins and myosin, a new study shows that formin-mediated polymerization is strongly inhibited upon the capture and pulling of actin filaments by myosin, a result that has broad implications for cellular mechanosensing.

remains incomplete. Here we present results using active micropost array detectors (AMPADS) to characterize the dynamical fluctuations and local rheology of cellular actomyosin networks in detail. AMPADS are poly(dimethylsiloxane) (PDMS) micropillar arrays with embedded magnetic nanowires that enable mechanical actuation of an adherent cell. We classified the cellassociated microposts based on their traction force, and identified two distinct populations, one containing posts coupled to stress fibers and the other containg posts coupled to the actomyosin cortex. Both groups show weak power law rheology, but we find that the fluctuations of stress fiber-associated posts are more active and highly anisotropic in comparison to the cortically-associated posts. Notably, the fluctuations of both populations are highly heterogenous and resemble a L evy process. Specifically, the mean-squared displacements of both types of posts display broadly distributed amplitudes and superdiffusive behavior. We find that the amplitude distribution is fat-tailed, and is dominated by a corresponding distribution of intermittent, large step-like displacements, resembling avalanches and earthquakes in physical systems. Our regular array of detectors also allows us to determine the spatial extent and average symmetry of the largest rearrangements in the cortex, which are both spatially and temporally complex, resembling those in plastic solids. These results suggest that the dynamics of the cellular actomyosin network resembles what is seen in soft matter systems such as sandpiles and foams, where constituent components self-organize into marginally stable, plastic networks, with significant implications for future models of the cortex. 2731-Pos Point Mutation of the Ice-Binding Site in Antifreeze Protein Modify the Cold Tolerance in Caenorhabditis Elegans