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Illuminating the dark phosphoproteome in Wnt signalling

Objectif

Phosphorylation has been considered as a fundamental on/off switch for protein function. However, many proteins harbour multiple phosphorylation sites arranged in dense clusters whose modification status is not reported in proteomic datasets due to technical limitations. Thus, multisite phosphorylation of dense clusters represents the dark matter of the phosphoproteome, since the very basic descriptors of function, regulation, and biology remain poorly understood. The presence of many target sites next to each other, ranging from 10 to as many as 30, raises new mechanistic questions with regard to the order of phosphorylation, or lack thereof.
DarkPhospho aims to elucidate the mechanisms and functional implications of multiphosphorylation using a panel of CK1 enzymes and their substrates – components of the Wnt pathway – as a model system and experimental toolkit. The project will take a multidisciplinary approach, comprising: (1) biophysical analysis of CK1 multiphosphorylation using nuclear magnetic resonance (NMR) and mass spectrometry (MS); (2) global kinetic analyses (computational numerical modelling) describing the stochastic nature of multiphosphorylation; and (3) role of multiphosphorylation in the regulation of protein phase separation (confocal microscopy) and Wnt signal transduction (dual luciferase reporter assay).
This project challenges the view that stochasticity at the molecular level is simply noise. The main scientific outcome will be a unified framework to analyse the diverse data and elucidate the harnessing of stochasticity in multiphosphorylation. The multidisciplinary design of the project offers the fellow extensive opportunities to diversify his research skills through training, gain valuable inter-sectoral experience, and extend his collaborative network, in line with his professional development needs and career objectives towards academic tenure.

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Mots‑clés

Programme(s)

Thème(s)

Régime de financement

HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Appel à propositions

(s’ouvre dans une nouvelle fenêtre) HORIZON-WIDERA-2025-TALENTS-01

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Coordinateur

Masarykova univerzita
Contribution nette de l'UE
€ 191 918,16
Adresse
Zerotinovo namesti 9
601 77 Brno
Tchéquie
Région
Česko Jihovýchod Jihomoravský kraj
Type d’activité
Higher or Secondary Education Establishments
Liens
Coût total
Aucune donnée