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FRET analysis of the spatio-temporal activation of Rho-family GTPases in T lymphocytes


Rho-family GTPases are signalling molecules that play a critical role during thymocyte development and T cell activation. The biochemical pathways regulated by Rho GTPases in lymphocytes are not at all understood although their importance is clear.

Recently the generation of mice deficient in Rho-family GTPases, their regulators and effectors have revealed functions for these GTPases at multiple regulatory levels, including the coordination of cytoskeletal change, activation of kinase cascades, stimulation of calcium fluxes, and the induction of gene expression. Information about the activation of Rho-family GTPases in T cells has been mainly provided by biochemical experiments, which allow only a global measure of GTPase activation.

However, it remains largely unknown when and where Rho-family GTPases are activated during T cell activation induced by various stimuli. Here we propose to use fluorescent resonance energy transfer (FRET)-based sensors to report the spatio-temporal activation of Rho-family GT Pases in living T cells in response to diverse stimuli such as TCR triggering, integrin engagement or chemokine stimulation.

We aim to perform this both in wild-type T cells and in T cells deficient in various guanine nucleotide exchange factors (GEFs) that are believed to activate Rho-family GTPases. In summary, this project will provide a better understanding of when, where and how different GTPases are activated in T cells in response to different signalling receptors.

Call for proposal

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Funding Scheme

EIF - Marie Curie actions-Intra-European Fellowships


20 Park Crescent
United Kingdom