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Updated: Mar 16, 2026
Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
Published on: December 20, 2014
Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins
Igor Cervenka1, Jana Valnohova1, Ondrej Bernatik2
1Department of Experimental Biology, Faculty of Science, Masaryk University, 61 137 Brno, Czech Republic;
Abstract:
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 subunit-associated 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.
Insights
Dishevelled (DVL) protein regulates the cell cycle by controlling centrosome separation. DVL
Area of Science:
- Cell Biology
- Molecular Biology
- Signal Transduction
Background:
- Dishevelled (DVL) is a crucial scaffolding protein in Wnt signaling pathways.
- The centrosome is vital for cell division, regulating spindle formation and chromosome segregation.
Purpose of the Study:
- To investigate the role of Dishevelled (DVL) in regulating the centrosomal cycle.
- To elucidate the molecular mechanisms linking DVL to centrosome function and Wnt signaling.
Main Methods:
- Utilized novel phospho-specific antibodies to detect DVL phosphorylation.
- Investigated protein-protein interactions using co-immunoprecipitation and localization studies.
- Examined the effects of DVL manipulation on centrosome separation and spindle formation.
Main Results:
- The dishevelled and axin (DIX) domain of DVL is essential for its centrosomal localization.
- DVL accumulates at the centrosome during the cell cycle and interacts with NEK2 kinase.
- NEK2-mediated phosphorylation of DVL facilitates its dissociation from the centrosome, promoting linker protein removal and centrosomal separation.
Conclusions:
- DVL plays a critical role in regulating centrosome separation through its interaction with NEK2 and centrosomal linker proteins.
- This study reveals a novel crosstalk between Wnt signaling and centrosome biology.
- Dysregulation of DVL at the centrosome can lead to mitotic spindle defects.
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