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Microtubules as platforms for probing liquid–liquid phase separation in cells – application to RNA-binding proteins

Abstract : Liquid-liquid phase separation enables compartmentalization of biomolecules in cells, notably RNA and associated proteins in the nucleus. Besides having critical functions in RNA processing, there is a major interest in deciphering the molecular mechanisms of compartmentalization orchestrated by RNA-binding proteins such as TDP-43 (also known as TARDBP) and FUS because of their link to neuron diseases. However, tools for probing compartmentalization in cells are lacking. Here, we developed a method to analyze the mixing and demixing of two different phases in a cellular context. The principle is the following: RNA-binding proteins are confined on microtubules and quantitative parameters defining their spatial segregation are measured along the microtubule network. Through this approach, we found that four mRNA-binding proteins, HuR (also known as ELAVL1), G3BP1, TDP-43 and FUS form mRNA-rich liquid-like compartments on microtubules. TDP-43 is partly miscible with FUS but immiscible with either HuR or G3BP1. We also demonstrate that mRNA is essential to capture the mixing and demixing behavior of mRNA-binding proteins in cells. Taken together, we show that microtubules can be used as platforms to understand the mechanisms underlying liquid-liquid phase separation and their deregulation in human diseases.
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Submitted on : Friday, October 18, 2019 - 1:38:00 PM
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(2018) Microtubules as platfor...
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Alexandre Maucuer, Bénédicte Desforges, Vandana Joshi, Mirela Boca, Dmitry Kretov, et al.. Microtubules as platforms for probing liquid–liquid phase separation in cells – application to RNA-binding proteins. Journal of Cell Science, Company of Biologists, 2018, 131 (11), pp.jcs214692. ⟨10.1242/jcs.214692⟩. ⟨hal-02166855⟩



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