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

Bernatík, Ondřej; Šedová, Kateřina; Schille, Carolin; Ganji, Ranjani Sri; Červenka, Igor; Trantı́rek, Lukáš; Schambony, Alexandra; Zdráhal, Zbyněk; Bryja, Vı́tězslav

doi:10.1074/jbc.m114.590638

Cited by 58 publications

(77 citation statements)

References 52 publications

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“…In 32D cells, heterologous expression of FZD 2 , FZD 4 , or FZD 5 resulted in the formation of PS-DVL even in the absence of exogenous WNT stimulation. These findings indicate that expression of FZD is sufficient to induce phosphorylation of endogenous DVLs, consistent with previous observations for FZD 5 (44). Finally, WNT-3A, -4, and -5A demonstrated functional activity in the 32D/FZD cell system as shown by ␤-catenin stabilization and phospho-LRP6 (WNT-3A) and PS-DVL2/3 (WNT-3A, -4, and -5A).…”

Section: Discussionsupporting

confidence: 77%

Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs

Dijksterhuis

Baljinnyam

Stanger³

et al. 2015

Journal of Biological Chemistry

149

135

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Background: WNT-FZD specificity and signaling outcome have not been systematically mapped so far. Results: WNTs show selective binding to FZDs, and respective WNT-FZD pairs exert functional selectivity in different downstream signaling pathways. Conclusion: The WNT-FZD signaling system provides ligand-receptor selectivity. Significance: Understanding WNT-FZD selectivity is crucial for development of WNT pathway inhibitors and further understanding of molecular interactions within the WNT receptor signaling complex.

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Section: Discussionsupporting

confidence: 77%

Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs

Dijksterhuis

Baljinnyam

Stanger³

et al. 2015

Journal of Biological Chemistry

149

135

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“…S3). Phosphorylation at S643 was associated with homogenous cytoplasmic localization of DVL3 (27), and indeed NEK2, similar to casein kinase (CK)1e coexpression (28,29), triggered subcellular relocalization of DVL from punctate to even cytoplasmic distribution (SI Appendix, Fig. S2D).…”

Section: Resultsmentioning

confidence: 99%

“…The efforts to explain DVL's individual "fates" by single-point mutations were not, so far, successful. For example, NEK2 resembles in many aspects the best-described DVL kinase CK1e: both kinases are capable of inducing a dramatic DVL phosphorylation shift, overlap in many target residues (e.g., S643 and S280 in hDVL3) (27), and are capable of promoting even cytoplasmic localization of DVL, but only CK1e can induce downstream Wnt/β-catenin signaling. It is likely that only identification and functional characterization of complex phosphorylation barcodes of individual DVL subcellular pools will fully reconcile the issue.…”

Section: Discussionmentioning

confidence: 99%

“…Dual luciferase was performed according to manufacturer's instruction and as described previously (29). Immunoprecipitation protocol used was performed as described previously (27). Expanded protocol can be found in SI Appendix.…”

Section: Methodsmentioning

confidence: 99%

“…Immunoblotting and sample preparation was performed as previously described (44) and developed using either light-sensitive films (Agfa, GE Healthcare) or chemiluminescence documentation system FusionSL (Vilber-Lourmat). MS for posttranslational modifications was performed as previously described (27), for full experimental details, see SI Appendix. Antibodies used for immunoprecipitation are summarized in SI Appendix, Table S3.…”

Section: Methodsmentioning

confidence: 99%

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Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins

Červenka

Valnohová

Bernatík

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Dishevelled (DVL) is a key scaffolding protein and a branching point in Wnt signaling pathways. Here, we present conclusive evidence that DVL regulates the centrosomal cycle. We demonstrate that DVL dishevelled and axin (DIX) domain, but not DIX domain-mediated multimerization, is essential for DVL's centrosomal localization. DVL accumulates during the cell cycle and associates with NIMA-related kinase 2 (NEK2), which is able to phosphorylate DVL at a multitude of residues, as detected by a set of novel phospho-specific antibodies. This creates interfaces for efficient binding to CDK5 regulatory subunitassociated protein 2 (CDK5RAP2) and centrosomal Nek2-associated protein 1 (C-NAP1), two proteins of the centrosomal linker. Displacement of DVL from the centrosome and its release into the cytoplasm on NEK2 phosphorylation is coupled to the removal of linker proteins, an event necessary for centrosomal separation and proper formation of the mitotic spindle. Lack of DVL prevents NEK2-controlled dissolution of loose centrosomal linker and subsequent centrosomal separation. Increased DVL levels, in contrast, sequester centrosomal NEK2 and mimic monopolar spindle defects induced by a dominant negative version of this kinase. Our study thus uncovers molecular crosstalk between centrosome and Wnt signaling.Wnt signaling | centrosome | Dishevelled | NEK2 | linker proteins

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CK1ε and p120‐catenin control Ror2 function in noncanonical Wnt signaling

et al. 2018

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Canonical and noncanonical Wnt pathways share some common elements but differ in the responses they evoke. Similar to Wnt ligands acting through the canonical pathway, Wnts that activate the noncanonical signaling, such as Wnt5a, promote Disheveled (Dvl) phosphorylation and its binding to the Frizzled (Fz) Wnt receptor complex. The protein kinase CK1ε is required for Dvl/Fz association in both canonical and noncanonical signaling. Here we show that differently to its binding to canonical Wnt receptor complex, CK1ε does not require p120‐catenin for the association with the Wnt5a co‐receptor Ror2. Wnt5a promotes the formation of the Ror2–Fz complex and enables the activation of Ror2‐bound CK1ε by Fz‐associated protein phosphatase 2A. Moreover, CK1ε also regulates Ror2 protein levels; CK1ε association stabilizes Ror2, which undergoes lysosomal‐dependent degradation in the absence of this kinase. Although p120‐catenin is not required for CK1ε association with Ror2, it also participates in this signaling pathway as p120‐catenin binds and maintains Ror2 at the plasma membrane; in p120‐depleted cells, Ror2 is rapidly internalized through a clathrin‐dependent mechanism. Accordingly, downregulation of p120‐catenin or CK1ε affects late responses to Wnt5a that are also sensitive to Ror2, such as SIAH2 transcription, cell invasion, or cortical actin polarization. Our results explain how CK1ε is activated by noncanonical Wnt and identify p120‐catenin and CK1ε as two critical factors controlling Ror2 function.

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

Cited by 58 publications

References 52 publications

Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs

Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs

Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins

CK1ε and p120‐catenin control Ror2 function in noncanonical Wnt signaling

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