J 2014

Functional Analysis of Dishevelled-3 Phosphorylation Identifies Distinct Mechanisms Driven by Casein Kinase 1 epsilon and Frizzled5

BERNATÍK, Ondřej, Kateřina ŠEDOVÁ, Carolin SCHILLE, Sri Ranjani GANJI, Igor ČERVENKA et. al.

Basic information

Original name

Functional Analysis of Dishevelled-3 Phosphorylation Identifies Distinct Mechanisms Driven by Casein Kinase 1 epsilon and Frizzled5

Authors

BERNATÍK, Ondřej (203 Czech Republic, belonging to the institution), Kateřina ŠEDOVÁ (203 Czech Republic, belonging to the institution), Carolin SCHILLE (276 Germany), Sri Ranjani GANJI (356 India, belonging to the institution), Igor ČERVENKA (703 Slovakia, belonging to the institution), Lukáš TRANTÍREK (203 Czech Republic, belonging to the institution), Alexandra SCHAMBONY (276 Germany), Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution) and Vítězslav BRYJA (203 Czech Republic, guarantor, belonging to the institution)

Edition

Journal of Biological Chemistry, Bethesda, USA, Amer. Soc. Biochem. Mol. Biol. 2014, 0021-9258

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 4.573

RIV identification code

RIV/00216224:14310/14:00074125

Organization unit

Faculty of Science

UT WoS

000341505000023

Keywords in English

PLANAR CELL POLARITY; BETA-CATENIN; WNT/BETA-CATENIN; PDZ DOMAIN; I-EPSILON; WNT PATHWAY; PROTEIN; DROSOPHILA; DVL; FAMILY

Tags

International impact, Reviewed
Změněno: 17/12/2014 12:57, prof. Mgr. Vítězslav Bryja, Ph.D.

Abstract

V originále

Background: Phosphorylation of Dishevelled (Dvl) by casein kinase 1E (CK1E) is a key event in Wnt signal transduction. Results: Dvl3 residues phosphorylated by CK1E were identified by proteomics and analyzed functionally. Conclusion: Individual phosphorylation events control different aspects of Dvl biology. Significance: CK1E and Fzd5, a Wnt receptor, act on Dvl via distinct mechanism, suggesting that CK1E is not directly downstream of Frizzled5. 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 1E (CK1E) was identified as the major kinase responsible for Wnt-induced Dvl3 phosphorylation. Currently it is not clear which Dvl residues are phosphorylated and what is the consequence of individual phosphorylation events. In the present study we employed mass spectrometry to analyze in a comprehensive way the phosphorylation of human Dvl3 induced by CK1E. Our analysis revealed >50 phosphorylation sites on Dvl3; only a minority of these sites was found dynamically induced after co-expression of CK1E, 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 CK1E 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 CK1E acts via phosphorylation of distinct residues as the activator as well as the shut-off signal of Wnt/-catenin signaling and (ii) suggest that CK1E acts on Dvl via different mechanism than Fzd5.

Links

ED1.1.00/02.0068, research and development project
Name: CEITEC - central european institute of technology
EE2.3.20.0042, research and development project
Name: Internacionalizace programu Strukturní biologie s důrazem na rozvoj nových směrů výzkumu
GA204/09/0498, research and development project
Name: Dynamika proteinů interagujících s Dishevelled a jejich význam pro Wnt signálování
Investor: Czech Science Foundation, Dynamics of proteins interacting with Dishevelled and their importance for Wnt signalling
LM2011020, research and development project
Name: CEITEC ? open access
Investor: Ministry of Education, Youth and Sports of the CR
MSM0021622430, plan (intention)
Name: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie
Investor: Ministry of Education, Youth and Sports of the CR, Functional and molecular characteristics of cancer and normal stem cells - identification of targets for novel therapeutics and therapeutic strategies