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.

Department of Experimental Biology Faculty of Science Masaryk University 61 137 Brno Czech Republic;

Department of Experimental Biology Faculty of Science Masaryk University 61 137 Brno Czech Republic; Department of Biochemistry and Biophysics Karolinska Institutet Stockholm 171 77 Sweden;

Department of Experimental Biology Faculty of Science Masaryk University 61 137 Brno Czech Republic; Institute of Biophysics Academy of Sciences of Czech Republic 61 200 Brno Czech Republic;

Department of Experimental Biology Faculty of Science Masaryk University 61 137 Brno Czech Republic; Receptor Biology and Signaling Department of Physiology and Pharmacology Karolinska Institutet 17 177 Stockholm Sweden;

Department of Histology and Embryology Faculty of Medicine Masaryk University 62 500 Brno Czech Republic;

Department of Pharmaceutical Sciences Leslie Dan Faculty of Pharmacy University of Toronto Toronto ON M5S 3M2 Canada;

Research Group Proteomics Central European Institute of Technology 62 500 Brno Czech Republic;

Research Group Proteomics Central European Institute of Technology 62 500 Brno Czech Republic; National Centre for Biomolecular Research Faculty of Science Masaryk University 625 00 Brno Czech Republic

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