1. Structural Biology of WNT Signalling
Supervisor : Konstantinos Tripsianes, Ph.D.
We apply structural biology methods in order to gain a mechanistic view of CK1ε action in the Wnt signaling pathways. CK1ε represents an attractive therapeutic target but currently two key steps in the CK1ε-mediated Wnt signal transduction are unclear: how CK1ε gets activated and/or engages target proteins in response to Wnt signal and how CK1ε phosphorylates its key substrate Dishevelled (DVL).
Our preliminary data suggest that we can efficiently apply methods of integrated structural biology to (i) probe the DVL conformational landscape using in vitro and in vivo FRET sensors coupled to SAXS and CryoEM, (ii) understand the (auto)phosphorylation regulatory mechanisms of CK1ε, (iii) analyse by NMR the functional consequences of DVL phosphorylation and (iv) monitor DVL phosphorylation by real-time NMR under controlled cellular conditions. The position is part of a multidisciplinary project that combines (i) cellular and molecular biology, (ii) proteomic analysis, (iii) biochemistry and structural biology, and received generous funding in a very competitive grant scheme.
Requirements on candidates:
- Biomolecular NMR
- Molecular Cell Biology
Recommended literature :
- Hanáková K., Bernatík O., Kravec M., Micka M., Kumar J., Harnoš J., Ovesná P., Paclíková P., Rádsetoulal M., Potěšil D., Tripsianes K., Čajánek L., Zdráhal Z.*, Bryja V.*, Comparative phosphorylation map of Dishevelled 3 links phospho-signatures to biological outputs. Cell Commun. Signal., 2019. 17: p. 170
- Harnoš J., Cañizal M.C.A., Jurásek M., Kumar J., Holler C., Schambony A., Hanáková K., Bernatík O., Zdráhal Z., Gömöryová K., Gybeľ T., Radaszkiewicz T.W., Kravec M., Trantírek L., Ryneš J., Dave Z., Fernández-Llamazares A.I., Vácha R., Tripsianes K., Hoffmann C., Bryja V.*, Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1. Nat. Commun., 2019. 10: p. 1804.
Keywords: Wnt signaling, Dishevelled, Casein kinase, interactions, conformations, phosphorylations, dynamics, allostery, NMR, X-ray crystallography, SAXS, FRET, cryo-EM, native MS, MS-HDX.