Abstract
The spatiotemporal organization of proteins within cells is essential for cell fate behavior. Although it is known that the cytoskeleton is vital for numerous cellular functions, it remains unclear how cytoskeletal activity can shape and control signaling pathways in space and time throughout the cell cytoplasm. Here we show that F-actin self-organization can trigger signaling pathways by engineering two novel properties of the microfilament self-organization: (1) the confinement of signaling proteins and (2) their scaffolding along actin polymers. Using in vitro reconstitutions of cellular functions, we found that both the confinement of nanoparticle-based signaling platforms powered by F-actin contractility and the scaffolding of engineered signaling proteins along actin microfilaments can drive a signaling switch. Using Ran-dependent microtubule nucleation, we found that F-actin dynamics promotes the robust assembly of microtubules. Our in vitro assay is a first step towards the development of novel bottom-up strategies to decipher the interplay between cytoskeleton spatial organization and signaling pathway activity.
Highlights
HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not
To examine how cytoplasmic confinement and partitioning can be mediated by cytoskeleton, we investigated the organization of F-actin filaments into dynamic contractile patterns using Metaphase II Xenopus egg extracts encapsulated within emulsion droplets
F-actin structures and dynamics drive numerous essential functions when they are assembled at the cortex of cells as well as within the cytoplasm
Summary
HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. Our in vitro assay is a first step towards the development of novel bottom-up strategies to decipher the interplay between cytoskeleton spatial organization and signaling pathway activity To achieve their numerous functionalities, cells have evolved a large variety of strategies to coordinate the spatial organization of intracellular components at multiple scales. Subcompartmentalization of signaling proteins mediated by the actin cytoskeleton has been proposed to regulate dendritic spines during neuronal plasticity[19] These recent works often describe mechanisms based on the interplay between F-actin operating at the membrane and signaling activity[14]. Recent studies highlight novel roles of cytoplasmic F-actin in various cellular functions[20,21,22,23,24,25,26], a general framework explaining how such cytoplasmic microfilaments may coordinate signaling pathways in space and time in the cytoplasm is still missing
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