Reconstitution of contractile actomyosin rings in vesicles.

One of the grand challenges of bottom-up synthetic biology is the development of minimal machineries for cell division. The mechanical transformation of large-scale compartments, such as Giant Unilamellar Vesicles (GUVs), requires the geometry-specific coordination of active elements, several orders of magnitude larger than the molecular scale. Of all cytoskeletal structures, large-scale actomyosin rings appear to be the most promising cellular elements to accomplish this task. Here, we have adopted advanced encapsulation methods to study bundled actin filaments in GUVs and compare our results with theoretical modeling. By changing few key parameters, actin polymerization can be differentiated to resemble various types of networks in living cells. Importantly, we find membrane binding to be crucial for the robust condensation into a single actin ring in spherical vesicles, as predicted by theoretical considerations. Upon force generation by ATP-driven myosin motors, these ring-like actin structures contract and locally constrict the vesicle, forming furrow-like deformations. On the other hand, cortex-like actin networks are shown to induce and stabilize deformations from spherical shapes.

Five-Site Model for Brownian Dynamics Simulations of a Molecular Walker in Three Dimensions.

Motor proteins play an important role in many biological processes and have inspired the development of synthetic analogues. Molecular walkers, such as kinesin, dynein, and myosin V, fulfill a diverse set of functions including transporting cargo along tracks, pulling molecules through membranes, and deforming fibers. The complexity of molecular motors and their environment makes it difficult to model the detailed dynamics of molecular walkers over long time scales. In this work, we present a simple, three-dimensional model for a molecular walker on a bead-spring substrate. The walker is represented by five spherically symmetric particles that interact through common intermolecular potentials and can be simulated efficiently in Brownian dynamics simulations. The movement of motor protein walkers entails energy conversion through ATP hydrolysis while artificial motors typically rely on a local conversion of energy supplied through external fields. We model energy conversion through rate equations for mechanochemical states that couple positional and chemical degrees of freedom and determine the walker conformation through interaction potential parameters. We perform Brownian dynamics simulations for two scenarios: In the first, the model walker transports cargo by walking on a substrate whose ends are fixed. In the second, a tethered motor pulls a mobile substrate chain against a variable force. We measure relative displacements and determine the effects of cargo size and retarding force on the efficiency of the walker. We find that, while the efficiency of our model walker is less than for the biological system, our simulations reproduce trends observed in single-molecule experiments on kinesin. In addition, the model and simulation method presented here can be readily adapted to biological and synthetic systems with multiple walkers.

Organization of associating or crosslinked actin filaments in confinement.

A key factor of actin cytoskeleton organization in cells is the interplay between the dynamical properties of actin filaments and cell geometry, which restricts, confines and directs their orientation. Crosslinking interactions among actin filaments, together with geometrical cues and regulatory proteins can give rise to contractile rings in dividing cells and actin rings in neurons. Motivated by recent in vitro experiments, in this work we performed computer simulations to study basic aspects of the interplay between confinement and attractive interactions between actin filaments. We used a spring-bead model and Brownian dynamics to simulate semiflexible actin filaments that polymerize in a confining sphere with a rate proportional to the monomer concentration. We model crosslinking, or attraction through the depletion interaction, implicitly as an attractive short-range potential between filament beads. In confining geometries smaller than the persistence length of actin filaments, we show rings can form by curving of filaments of length comparable to, or longer than the confinement diameter. Rings form for optimal ranges of attractive interactions that exist in between open bundles, irregular loops, aggregated, and unbundled morphologies. The probability of ring formation is promoted by attraction to the confining sphere boundary and decreases for large radii and initial monomer concentrations, in agreement with prior experimental data. The model reproduces ring formation along the flat plane of oblate ellipsoids.