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

0520259 - BFÚ 2020 RIV GB eng J - Článek v odborném periodiku
Harnoš, J. - Canizal, M.C.A. - Jurasek, M. - Kumar, J. - Holler, C. - Schambony, A. - Hanakova, K. - Bernatík, O. - Zdráhal, Z. - Gomoryova, K. - Gybel, T. - Radaszkiewicz, T.W. - Kravec, M. - Trantírek, Lukáš - Ryneš, J. - Dave, Z. - Fernandez-Llamazares, A.I. - Vácha, R. - Tripsianes, K. - Hoffmann, C. - Bryja, Vítězslav
Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1.
Nature Communications. Roč. 10, APR 2019 (2019), č. článku 1804. E-ISSN 2041-1723
Grant CEP: GA MŠMT EF15_003/0000477
Institucionální podpora: RVO:68081707
Klíčová slova: particle mesh ewald * beta-catenin * wnt/beta-catenin * dix domain
Obor OECD: Biochemical research methods
Impakt faktor: 12.121, rok: 2019
Způsob publikování: Open access
https://www.nature.com/articles/s41467-019-09651-7.pdf

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.
Trvalý link: http://hdl.handle.net/11104/0304964