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Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1
J. Harnoš, MCA. Cañizal, M. Jurásek, J. Kumar, C. Holler, A. Schambony, K. Hanáková, O. Bernatík, Z. Zdráhal, K. Gömöryová, T. Gybeľ, TW. Radaszkiewicz, M. Kravec, L. Trantírek, J. Ryneš, Z. Dave, AI. Fernández-Llamazares, R. Vácha, K....
Jazyk angličtina Země Anglie, Velká Británie
Typ dokumentu časopisecké články, práce podpořená grantem
Grantová podpora
NV15-34405A
MZ0
CEP - Centrální evidence projektů
Digitální knihovna NLK
Plný text - Článek
Zdroj
NLK
Directory of Open Access Journals
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od 2010-12-01
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od 2015-01-01
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od 2015-01-01
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od 2012-11-01
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od 2010-01-01
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od 2010
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od 2010-12-01
- MeSH
- analýza jednotlivých buněk metody MeSH
- biosenzitivní techniky MeSH
- enzymatické testy metody MeSH
- fluorescenční mikroskopie metody MeSH
- fosforylace fyziologie MeSH
- frizzled receptory metabolismus MeSH
- genový knockout MeSH
- HEK293 buňky MeSH
- kaseinkinasa Iepsilon genetika metabolismus MeSH
- lidé MeSH
- mutageneze cílená MeSH
- oocyty MeSH
- PDZ domény fyziologie MeSH
- protein dishevelled genetika metabolismus MeSH
- rezonanční přenos fluorescenční energie MeSH
- signální dráha Wnt fyziologie MeSH
- simulace molekulární dynamiky MeSH
- Xenopus laevis MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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.
CEITEC Central European Institute of Technology Masaryk University Brno 62500 Czech Republic
Department of Experimental Biology Faculty of Science Masaryk University Brno 62500 Czech Republic
Citace poskytuje Crossref.org
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- $a Harnoš, Jakub $u 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.
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- $a Dishevelled-3 conformation dynamics analyzed by FRET-based biosensors reveals a key role of casein kinase 1 / $c J. Harnoš, MCA. Cañizal, M. Jurásek, J. Kumar, C. Holler, A. Schambony, K. Hanáková, O. Bernatík, Z. Zdráhal, K. Gömöryová, T. Gybeľ, TW. Radaszkiewicz, M. Kravec, L. Trantírek, J. Ryneš, Z. Dave, AI. Fernández-Llamazares, R. Vácha, K. Tripsianes, C. Hoffmann, V. Bryja,
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- $a Cañizal, Maria Consuelo Alonso $u 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.
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