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Assessment of Dictyostelium discoideum Response to Acute Mechanical Stimulation
Published on: November 9, 2017
Oscillatory dynamics of Rac1 activity in Dictyostelium discoideum amoebae
Marko Šoštar1, Maja Marinović1, Vedrana Filić1
1Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Abstract:
Small GTPases of the Rho family play a central role in the regulation of cell motility by controlling the remodeling of the actin cytoskeleton. In the amoeboid cells of Dictyostelium discoideum, the active form of the Rho GTPase Rac1 regulates actin polymerases at the leading edge and actin filament bundling proteins at the posterior cortex of polarized cells. We monitored the spatiotemporal dynamics of Rac1 and its effector DGAP1 in vegetative amoebae using specific fluorescent probes. We observed that plasma membrane domains enriched in active Rac1 not only exhibited stable polarization, but also showed rotations and oscillations, whereas DGAP1 was depleted from these regions. To simulate the observed dynamics of the two proteins, we developed a mass-conserving reaction-diffusion model based on the circulation of Rac1 between the membrane and the cytoplasm coupled with its activation by GEFs, deactivation by GAPs and interaction with DGAP1. Our theoretical model accurately reproduced the experimentally observed dynamic patterns, including the predominant anti-correlation between active Rac1 and DGAP1. Significantly, the model predicted a new colocalization regime of these two proteins in polarized cells, which we confirmed experimentally. In summary, our results improve the understanding of Rac1 dynamics and reveal how the occurrence and transitions between different regimes depend on biochemical reaction rates, protein levels and cell size. This study not only expands our knowledge of the behavior of Rac1 GTPases in D. discoideum amoebae but also demonstrates how specific modes of interaction between Rac1 and its effector DGAP1 lead to their counterintuitively anti-correlated dynamics.
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