title: Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins.
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.
authors.affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61 137 Brno, Czech Republic;
authors.forename: Igor
authors.initials: I
authors.lastname: Cervenka
authors.affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61 137 Brno, Czech Republic;
authors.forename: Jana
authors.initials: J
authors.lastname: Valnohova
authors.affiliation: 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;
authors.forename: Ondrej
authors.initials: O
authors.lastname: Bernatik
authors.affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61 137 Brno, Czech Republic;
authors.forename: Jakub
authors.initials: J
authors.lastname: Harnos
authors.affiliation: Department of Experimental Biology, Faculty of Science, Masaryk University, 61 137 Brno, Czech Republic;
authors.forename: Matej
authors.initials: M
authors.lastname: Radsetoulal
authors.affiliation: Research Group Proteomics, Central European Institute of Technology, 62 500 Brno, Czech Republic;
authors.forename: Katerina
authors.initials: K
authors.lastname: Sedova
authors.affiliation: Research Group Proteomics, Central European Institute of Technology, 62 500 Brno, Czech Republic;
authors.forename: Katerina
authors.initials: K
authors.lastname: Hanakova
authors.affiliation: Research Group Proteomics, Central European Institute of Technology, 62 500 Brno, Czech Republic;
authors.forename: David
authors.initials: D
authors.lastname: Potesil
authors.affiliation: Department of Histology and Embryology, Faculty of Medicine, Masaryk University, 62 500 Brno, Czech Republic;
authors.forename: Miroslava
authors.initials: M
authors.lastname: Sedlackova
authors.affiliation: 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;
authors.forename: Alena
authors.initials: A
authors.lastname: Salasova
authors.affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON M5S 3M2, Canada;
authors.forename: Zachary
authors.initials: Z
authors.lastname: Steinhart
authors.affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON M5S 3M2, Canada;
authors.forename: Stephane
authors.initials: S
authors.lastname: Angers
authors.affiliation: 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;
authors.forename: Gunnar
authors.initials: G
authors.lastname: Schulte
authors.affiliation: Department of Histology and Embryology, Faculty of Medicine, Masaryk University, 62 500 Brno, Czech Republic;
authors.forename: Ales
authors.initials: A
authors.lastname: Hampl
authors.affiliation: 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.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Zdrahal
authors.affiliation: 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; bryja@sci.muni.cz.
authors.forename: Vitezslav
authors.initials: V
authors.lastname: Bryja
mesh_terms: D001324:Autoantigens / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D018385:Centrosome / Q000378:metabolism*; D000072261:Dishevelled Proteins / Q000502:physiology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D000072379:NIMA-Related Kinases / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010766:Phosphorylation; D060449:Wnt Signaling Pathway
chemical_list: D001324:Autoantigens; C405929:CDK5RAP2 protein, human; D018797:Cell Cycle Proteins; C111329:CEP250 protein, human; D000072261:Dishevelled Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; C089441:NEK2 protein, human; D000072379:NIMA-Related Kinases
keywords: Dishevelled; NEK2; Wnt signaling; centrosome; linker proteins