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bioRxiv · 10.1101/2024.11.04.621887

Actin turnover and myosin contractility determine emergent Thickness Robustness of Cell-Mimicking Cortex

Gat, S.·Amit, Y.·Livne, G.·Sanchez, D. S.·Porat, N.·Yochelis, A.·Gov, N.·Bernheim-Groswasser, A.
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Abstract

Cell shape deformation in eukaryotes, is primarily determined by the actin cortex, a thin network of actin filaments and myosin motors. This network is attached to the plasma membrane endowing cells with their mechanical stability and structural integrity. Since inside the cells hundreds of proteins are associated with the actin cortex, identifying the roles of the individual components and the mechanisms maintaining a functional cortex is highly challenging. Here, using a minimal set of components, we succeeded in recreating long-lasting dynamic treadmilling in an artificial actomyosin cortex on a bilayer membrane with cell-mimicking characteristics, both with and without myosin-II motors, replicating features observed in cells, such as myosin-induced increased cortical thickness and stress-dependent dynamics. We mechanistically reveal how treadmilling is regulated by actin network disassembly factors and myosin contractility, and the emergence of robustness of the cortical thickness to concentration fluctuations in cytoskeletal components. The robustness and responsiveness of the actin cortex which we found has a fundamental significance for the ability of Eukaryotic cells to maintain the mechanical integrity of their membrane, despite concentration fluctuation of the cytoskeleton components. Unlike complex biological cells, this system enables high-resolution and systematic studies of cortical dynamics under controlled conditions. It thus provides a cell-mimicking artificial system for designing interventions to modify the cellular cortex to improve our understanding of the basic mechanisms driving fundamental biological processes as well as for potential medical applications. SignificanceWe report a breakthrough in understanding the actin cortex, a key structure governing cell shape, stability, and motility. By reconstructing in-vitro a treadmilling actomyosin cortex on a lipid bilayer using minimal components, we replicate essential cellular features. This system reveals how cortical thickness is maintained despite concentration fluctuations, highlighting an actin turnover mechanism that buffers variability. The inclusion of myosin motors is found to modify the cortical dynamics, mimicking cellular properties such as the increased thickness and stress-dependent dynamics. Unlike complex living cells, this artificial system enables systematic studies of cortical behavior, with fine control over the components and high-resolution imaging. Our artificial system provides a platform for designing interventions to modify cortical dynamics, with broad implications for biology and medicine. One-Sentence SummaryIn-vitro recreation of a cell-like, membrane-bound cortical actin skeleton composed of a minimal set of building-blocks with continuous turnover and emergent robustness.

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Gat, S., Amit, Y., Livne, G., Sanchez, D. S., Porat, N., Yochelis, A., Gov, N., Bernheim-Groswasser, A.. 2024-11-06. Actin turnover and myosin contractility determine emergent Thickness Robustness of Cell-Mimicking Cortex. https://doi.org/10.1101/2024.11.04.621887

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