A Novel Stochastic Simulation Approach Enables Exploration Of Mechanisms For Regulating Polarity Site Movement

Author summary Cells localize polarity molecules in a small region of the plasma membrane forming a polarity cluster that directs functions such as migration, reproduction, and growth. Guided by external signals, these clusters move across the membrane allowing cells to reorient growth or motion. The polarity molecules continuously and randomly shuttle between the cluster and the cell cytosol and, as a result, the number and distribution of molecules at the cluster constantly changes. Here we present an improved stochastic simulation algorithm to investigate how such molecular-scale fluctuations induce cluster movement across the cell membrane. Unexpectedly, cluster mobility does not correlate with variations in total molecule abundance within the cluster, but rather with changes in the spatial distribution of molecules that form the cluster. Cluster motion is faster when polarity molecules are scarce and when they shuttle rapidly between the cluster and the cytosol. Our results suggest that cells control cluster mobility by regulating the abundance of polarity molecules and biochemical reactions that affect the time molecules spend at the cluster. We provide insights into how cells harness random molecular behavior to perform functions important for survival, such as detecting the direction of external signals.Cells polarize their movement or growth toward external directional cues in many different contexts. For example, budding yeast cells grow toward potential mating partners in response to pheromone gradients. Directed growth is controlled by polarity factors that assemble into clusters at the cell membrane. The clusters assemble, disassemble, and move between different regions of the membrane before eventually forming a stable polarity site directed toward the pheromone source. Pathways that regulate clustering have been identified but the molecular mechanisms that regulate cluster mobility are not well understood. To gain insight into the contribution of chemical noise to cluster behavior we simulated clustering within the reaction-diffusion master equation (RDME) framework to account for molecular-level fluctuations. RDME simulations are a computationally efficient approximation, but their results can diverge from the underlying microscopic dynamics. We implemented novel concentration-dependent rate constants that improved the accuracy of RDME-based simulations of cluster behavior, allowing us to efficiently investigate how cluster dynamics might be regulated. Molecular noise was effective in relocating clusters when the clusters contained low numbers of limiting polarity factors, and when Cdc42, the central polarity regulator, exhibited short dwell times at the polarity site. Cluster stabilization occurred when abundances or binding rates were altered to either lengthen dwell times or increase the number of polarity molecules in the cluster. We validated key results using full 3D particle-based simulations. Understanding the mechanisms cells use to regulate the dynamics of polarity clusters should provide insights into how cells dynamically track external directional cues.