Functional analysis of dishevelled-3 phosphorylation identifies distinct mechanisms driven by casein kinase 1ϵ and frizzled5.

Beitrag in einer Fachzeitschrift
(Originalarbeit)


Details zur Publikation

Autorinnen und Autoren: Bernatik O, Sedova K, Schille C, Ganji RS, Cervenka I, Trantirek L, Schambony A, Zdrahal Z, Bryja V
Zeitschrift: Journal of Biological Chemistry
Jahr der Veröffentlichung: 2014
Band: 289
Heftnummer: 34
Seitenbereich: 23520-23533
ISSN: 0021-9258


Abstract


Dishevelled-3 (Dvl3), a key component of the Wnt signaling pathways, acts downstream of Frizzled (Fzd) receptors and gets heavily phosphorylated in response to pathway activation by Wnt ligands. Casein kinase 1ε (CK1ε) was identified as the major kinase responsible for Wnt-induced Dvl3 phosphorylation. Currently it is not clear which Dvl residues are phosphorylatedandwhatis theconsequenceof individualphosphorylation events. In the present study we employed mass spectrometry to analyze in a comprehensive way the phosphorylation of human Dvl3 induced by CK1ε. Our analysis revealed >50 phosphorylation sites on Dvl3; only a minority of these sites was found dynamically induced after co-expression of CK1ε, and surprisingly, phosphorylation of one cluster of modified residues was down-regulated. Dynamically phosphorylated sites were analyzed functionally. Mutations within PDZ domain (S280A and S311A) reduced the ability of Dvl3 to activate TCF/ LEF (T-cell factor/lymphoid enhancer factor)-driven transcription and induce secondary axis in Xenopus embryos. In contrast, mutations of clustered Ser/Thr in the Dvl3 C terminus prevented ability of CK1ε to induce electrophoretic mobility shift of Dvl3 and its even subcellular localization. Surprisingly, mobility shift and subcellular localization changes induced by Fzd5, a Wnt receptor, were in all these mutants indistinguishable from wild type Dvl3. In summary, our data on the molecular level (i) support previous the assumption that CK1ε acts via phosphorylation of distinct residues as the activator as well as the shut-off signal of Wnt/β-catenin signaling and (ii) suggest that CK1ε acts on Dvl via different mechanism than Fzd5. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.



FAU-Autorinnen und Autoren / FAU-Herausgeberinnen und Herausgeber

Schambony, Alexandra Prof. Dr.
Lehrstuhl für Entwicklungsbiologie
Schille, Carolin
Lehrstuhl für Entwicklungsbiologie


Einrichtungen weiterer Autorinnen und Autoren

Masaryk University


Zitierweisen

APA:
Bernatik, O., Sedova, K., Schille, C., Ganji, R.S., Cervenka, I., Trantirek, L.,... Bryja, V. (2014). Functional analysis of dishevelled-3 phosphorylation identifies distinct mechanisms driven by casein kinase 1ϵ and frizzled5. Journal of Biological Chemistry, 289(34), 23520-23533. https://dx.doi.org/10.1074/jbc.M114.590638

MLA:
Bernatik, Ondrej, et al. "Functional analysis of dishevelled-3 phosphorylation identifies distinct mechanisms driven by casein kinase 1ϵ and frizzled5." Journal of Biological Chemistry 289.34 (2014): 23520-23533.

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