Functional consequences of Wnt-induced dishevelled 2 phosphorylation in canonical and noncanonical Wnt signaling

J Biol Chem. 2013 Mar 29;288(13):9428-37. doi: 10.1074/jbc.M112.448480. Epub 2013 Feb 8.


Dishevelled (Dvl) proteins are intracellular effectors of Wnt signaling that have essential roles in both canonical and noncanonical Wnt pathways. It has long been known that Wnts stimulate Dvl phosphorylation, but relatively little is known about its functional significance. We have previously reported that both Wnt3a and Wnt5a induce Dvl2 phosphorylation that is associated with an electrophoretic mobility shift and loss of recognition by monoclonal antibody 10B5. In the present study, we mapped the 10B5 epitope to a 16-amino acid segment of human Dvl2 (residues 594-609) that contains four Ser/Thr residues. Alanine substitution of these residues (P4m) eliminated the mobility shift induced by either Wnt3a or Wnt5a. The Dvl2 P4m mutant showed a modest increase in canonical Wnt/β-catenin signaling activity relative to wild type. Consistent with this finding, Dvl2 4Pm preferentially localized to cytoplasmic puncta. In contrast to wild-type Dvl2, however, the P4m mutant was unable to rescue Wnt3a-dependent neurite outgrowth in TC-32 cells following suppression of endogenous Dvl2/3. Earlier work has implicated casein kinase 1δ/ε as responsible for the Dvl mobility shift, and a CK1δ in vitro kinase assay confirmed that Ser(594), Thr(595), and Ser(597) of Dvl2 are CK1 targets. Alanine substitution of these three residues was sufficient to abrogate the Wnt-dependent mobility shift. Thus, we have identified a cluster of Ser/Thr residues in the C-terminal domain of Dvl2 that are Wnt-induced phosphorylation (WIP) sites. Our results indicate that phosphorylation at the WIP sites reduces Dvl accumulation in puncta and attenuates β-catenin signaling, whereas it enables noncanonical signaling that is required for neurite outgrowth.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Adaptor Proteins, Signal Transducing / physiology*
  • Alanine / chemistry
  • Animals
  • Culture Media, Conditioned
  • Dishevelled Proteins
  • Gene Expression Regulation*
  • HEK293 Cells
  • Humans
  • Mutation
  • Phosphoproteins / metabolism
  • Phosphoproteins / physiology*
  • Phosphorylation
  • Proto-Oncogene Proteins / metabolism
  • RNA, Small Interfering / metabolism
  • Rats
  • Serine / chemistry
  • Signal Transduction
  • Threonine / chemistry
  • Wnt Proteins / metabolism*
  • Wnt-5a Protein
  • Wnt3A Protein / metabolism


  • Adaptor Proteins, Signal Transducing
  • Culture Media, Conditioned
  • DVL2 protein, human
  • Dishevelled Proteins
  • Phosphoproteins
  • Proto-Oncogene Proteins
  • RNA, Small Interfering
  • WNT3A protein, human
  • WNT5A protein, human
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3A Protein
  • Wnt5a protein, rat
  • Threonine
  • Serine
  • Alanine