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

Autor:	Harnoš J; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic.; Department of Cell, Developmental & Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Cañizal MCA; Department of Pharmacology and Toxicology, University of Würzburg, Würzburg, 97078, Germany.; Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Würzburg, 97078, Germany.; Institute for Molecular Cell Biology, CMB-Center for Molecular Biomedicine, University Hospital Jena, Friedrich Schiller University Jena, Jena, 07745, Germany., Jurásek M; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Kumar J; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic., Holler C; Max Planck Institute for the Science of Light, Erlangen, 91058, Germany.; Biology Department, Developmental Biology, Friedrich-Alexander University Erlangen-Nüremberg, Erlangen, 91058, Germany., Schambony A; Max Planck Institute for the Science of Light, Erlangen, 91058, Germany.; Biology Department, Developmental Biology, Friedrich-Alexander University Erlangen-Nüremberg, Erlangen, 91058, Germany., Hanáková K; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Bernatík O; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Zdráhal Z; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Gömöryová K; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Gybeľ T; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Radaszkiewicz TW; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Kravec M; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Trantírek L; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.; Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Brno, 612 65, Czech Republic., Ryneš J; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic., Dave Z; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Fernández-Llamazares AI; Pepscan Therapeutics B.V., Lelystad, 8243, The Netherlands., Vácha R; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.; National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic., Tripsianes K; CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic., Hoffmann C; Department of Pharmacology and Toxicology, University of Würzburg, Würzburg, 97078, Germany.; Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Würzburg, 97078, Germany.; Institute for Molecular Cell Biology, CMB-Center for Molecular Biomedicine, University Hospital Jena, Friedrich Schiller University Jena, Jena, 07745, Germany., Bryja V; Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic. bryja@sci.muni.cz.; Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Brno, 612 65, Czech Republic. bryja@sci.muni.cz.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2019 Apr 18; Vol. 10 (1), pp. 1804. Date of Electronic Publication: 2019 Apr 18.
DOI:	10.1038/s41467-019-09651-7
Abstrakt:	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.
Databáze:	MEDLINE
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