Wnt stimulus leads to Casein kinase (CK) 1 δ/ε-mediated Dishevelled (DVL) phosphorylation that was investigated by both NMR spectroscopy and phosphoproteomics. One of the applied methods was MALDI-MS. DVL phosphorylation results in the accumulation of negative charge and when the charge threshold is reached, DVL conformation is altered, as revealed by NMR spectroscopy. Furthermore, proximity interactomics revealed that DVL dissociates from Frizzled (FZD) receptors after DVL conformational switch and transduces the signal downstream. Created using BioRender. Gybel, T. (2026) https://BioRender.com/tremyul. All the NMR, phosphoproteomics and proximity interactomics analyses were executed at the CIISB core facilities.
Laboratory of Konstantinos Tripsianes & Vítězslav Bryja
Significance
Laboratory of Konstantinos Tripsianes (CEITEC MU) and Vítězslav Bryja (Faculty of Science MU) conducted a collaborative research to understand the mechanism of Wnt signal transduction from the receptor at the plasma membrane to the cytosolic downstream components. They focused on cytoplasmic protein Dishevelled (DVL) that becomes phosphorylated when Wnt pathway is activated.
Collaboration of both research groups enabled to study DVL phosphorylation both in vitro and in the cellular environment. In vitro data revealed a charge-induced conformational change of DVL when the charge threshold is reached, after a certain number of phosphate groups is deposited on DVL. Furthermore, researchers were able to link these findings with the functional cellular data and revealed that DVL dissociates from the receptor after the conformational change.
The use of Josef Dadok National NMR Centre and CEITEC Proteomics Core Facility of CIISB, Instruct-CZ Centre, supported by MEYS CR (LM2023042, CZ.02.01.01/00/23_015/0008175, e-INFRA CZ (ID:90254)) was crucial for the experimental success of this study.
The results were published in Science Advances. https://www.science.org/doi/10.1126/sciadv.aed8899