Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1

• Autoři: HARNOŠ Jakub; CANIZAL M.C.A.; JURÁSEK Miroslav; KUMAR Jitender; HOLLER Cornelia; SCHAMBONY Alexandra; HANÁKOVÁ Kateřina; BERNATÍK Ondřej; ZDRÁHAL Zbyněk; GÖMÖRYOVÁ Kristína; GYBEĽ Tomáš; RADASZKIEWICZ Tomasz Witold; KRAVEC Marek; TRANTÍREK Lukáš; RYNEŠ Jan; DAVE Zankruti; FERNANDEZ-LLAMAZARES Ana Iris; VÁCHA Robert; TRIPSIANES Konstantinos; HOFFMANN Carsten; BRYJA Vítězslav
• Rok publikování: 2019
• Druh: Článek v odborném periodiku
• Časopis / Zdroj: Nature Communications
• Fakulta / Pracoviště MU: Přírodovědecká fakulta
• www: Full Text
• Doi: http://dx.doi.org/10.1038/s41467-019-09651-7
• Klíčová slova: PARTICLE MESH EWALD; BETA-CATENIN; WNT/BETA-CATENIN; DIX DOMAIN; MOLECULAR-DYNAMICS; STRUCTURAL BASIS; PDZ DOMAIN; DEP DOMAIN; G-PROTEIN; WNT
• Popis: Dishevelled (DVL) is the key component of the Wnt signaling pathway. Currently, DVL conformational dynamics under native conditions is unknown. To overcome this limitation, we develop the Fluorescein Arsenical Hairpin Binder- (FlAsH-) based FRET in vivo approach to study DVL conformation in living cells. Using this single-cell FRET approach, we demonstrate that (i) Wnt ligands induce open DVL conformation, (ii) DVL variants that are predominantly open, show more even subcellular localization and more efficient membrane recruitment by Frizzled (FZD) and (iii) Casein kinase 1 epsilon (CK1 epsilon) has a key regulatory function in DVL conformational dynamics. In silico modeling and in vitro biophysical methods explain how CK1 epsilon-specific phosphorylation events control DVL conformations via modulation of the PDZ domain and its interaction with DVL C-terminus. In summary, our study describes an experimental tool for DVL conformational sampling in living cells and elucidates the essential regulatory role of CK1 epsilon in DVL conformational dynamics.

Související projekty

• WNT-mediated signal relay in stem cells and oncogenesis - from basic biology to applications
• Dishevelled - identifikace základních koordinát
• Identifikace nových možností léčby chronické myeloidní leukémie pomocí systematické analýzy interaktomu proteinu BCR-ABL
• CERIT Scientific Cloud
• Czech Infrastructure for Integrative Structural Biology
• CEITEC 2020
• Computational chemistry for Wnt signaling pathway
• Odhalení tajemství signální dráhy WNT analýzou struktury proteinu Dishevelled
• Pushing the limits in automated NMR structure determination using a single 4D NOESY spectrum and machine learning methods
• Studium konformační dynamiky proteinu Dishevelled 3 pomocí FRET sensorů: objev klíčové role Kasein kinázy 1 (CK1).
• Preklinická progrese nových organických sloučenin s cílenou biologickou aktivitou
• STRUCTURAL BIOLOGY – NEXT GENERATION: from isolated molecules to cells, from cells to tissues.