Cargando…

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

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 c...

Descripción completa

Detalles Bibliográficos
Autores principales: Harnoš, Jakub, Cañizal, Maria Consuelo Alonso, 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, Fernández-Llamazares, Ana Iris, Vácha, Robert, Tripsianes, Konstantinos, Hoffmann, Carsten, Bryja, Vítězslav
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6472409/
https://www.ncbi.nlm.nih.gov/pubmed/31000703
http://dx.doi.org/10.1038/s41467-019-09651-7
Descripción
Sumario: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 ɛ (CK1ɛ) has a key regulatory function in DVL conformational dynamics. In silico modeling and in vitro biophysical methods explain how CK1ɛ-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 CK1ɛ in DVL conformational dynamics.