title: Functional consequences of Wnt-induced dishevelled 2 phosphorylation in canonical and noncanonical Wnt signaling.
abstract: 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.
authors: Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY 10065, USA.
authors: José M
authors: JM
authors: González-Sancho
authors: Yoshimi Endo
authors: YE
authors: Greer
authors: Cristina L
authors: CL
authors: Abrahams
authors: Yutaka
authors: Y
authors: Takigawa
authors: Bolormaa
authors: B
authors: Baljinnyam
authors: Kyung Ho
authors: KH
authors: Lee
authors: Kyung S
authors: KS
authors: Lee
authors: Jeffrey S
authors: JS
authors: Rubin
authors: Anthony M C
authors: AM
authors: Brown
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism / Q000502:physiology*; D000409:Alanine / Q000737:chemistry; D000818:Animals; D017077:Culture Media, Conditioned; D000072261:Dishevelled Proteins; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D010750:Phosphoproteins / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction; D013912:Threonine / Q000737:chemistry; D051153:Wnt Proteins / Q000378:metabolism*; D000071818:Wnt-5a Protein; D060509:Wnt3A Protein / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D017077:Culture Media, Conditioned; C000606581:DVL2 protein, human; D000072261:Dishevelled Proteins; D010750:Phosphoproteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; C555510:WNT3A protein, human; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D060509:Wnt3A Protein; C491837:Wnt5a protein, rat; D013912:Threonine; D012694:Serine; D000409:Alanine
references: Nat Rev Genet. 2007 Feb;8(2):126-38
references: 17230199
references: Cell. 2010 Dec 23;143(7):1136-48
references: 21183076
references: Mol Cell Biol. 2008 Apr;28(7):2368-79
references: 18212053
references: J Neurochem. 2005 Mar;92(6):1550-3
references: 15748172
references: Cell Signal. 2010 May;22(5):717-27
references: 20006983
references: Dev Biol. 2011 Dec 1;360(1):132-42
references: 21963539
references: Nat Struct Mol Biol. 2007 Jun;14(6):484-92
references: 17529994
references: Science. 2007 Jun 15;316(5831):1619-22
references: 17569865
references: Cell. 2006 Nov 3;127(3):469-80
references: 17081971
references: Dev Biol. 1994 Nov;166(1):73-86
references: 7958461
references: Cell. 2007 Dec 28;131(7):1378
references: 18160045
references: Cell Signal. 2007 Mar;19(3):610-6
references: 17027228
references: PLoS One. 2012;7(1):e29455
references: 22238613
references: Genes Dev. 1994 Jan;8(1):118-30
references: 8288125
references: Dev Cell. 2009 Jul;17(1):9-26
references: 19619488
references: Methods Mol Biol. 2008;468:131-44
references: 19099251
references: Oncogene. 2010 Jan 7;29(1):34-44
references: 19802008
references: J Biol Chem. 2011 Mar 25;286(12):10396-410
references: 21285348
references: Oncogene. 2009 Sep 10;28(36):3197-208
references: 19561643
references: Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12548-52
references: 10535959
references: J Biol Chem. 1999 Jul 23;274(30):21464-70
references: 10409711
references: Cell Signal. 2005 Jun;17(6):675-89
references: 15722192
references: Br J Cancer. 2009 Jul 21;101(2):209-14
references: 19603030
references: Trends Cell Biol. 2010 Jun;20(6):346-54
references: 20359892
references: J Cell Biol. 2001 Sep 3;154(5):983-93
references: 11524435
references: Cell. 2001 Dec 28;107(7):843-54
references: 11779461
references: Genomics. 1997 Jun 1;42(2):302-10
references: 9192851
references: FEBS J. 2006 Oct;273(20):4594-602
references: 16965538
references: J Biol Chem. 2005 Jan 7;280(1):777-86
references: 15509575
references: Nat Rev Cancer. 2008 May;8(5):387-98
references: 18432252
references: Curr Biol. 1996 Oct 1;6(10):1302-6
references: 8939578
references: Cell. 2002 Mar 22;108(6):837-47
references: 11955436
references: Cell Growth Differ. 1997 Dec;8(12):1349-58
references: 9419423
references: J Cell Sci. 2005 Nov 15;118(Pt 22):5269-77
references: 16263762
references: J Biol Chem. 2000 Feb 11;275(6):4374-82
references: 10660608
references: J Cell Biol. 2011 Mar 21;192(6):993-1004
references: 21422228
references: J Cell Sci. 2007 Feb 15;120(Pt 4):586-95
references: 17244647
references: Dev Biol. 2003 Jan 1;253(1):1-17
references: 12490194
references: Int Rev Cell Mol Biol. 2011;291:21-71
references: 22017973
references: Nature. 1994 Mar 24;368(6469):342-4
references: 8127369
references: Mol Cell Biol. 2004 Jun;24(11):4757-68
references: 15143170
references: J Cell Sci. 2007 Jul 15;120(Pt 14):2402-12
references: 17606995
references: Nature. 1999 Sep 23;401(6751):345-50
references: 10517632
references: EMBO J. 2010 Oct 20;29(20):3470-83
references: 20823832
references: Dev Cell. 2003 Sep;5(3):367-77
references: 12967557
references: J Biol Chem. 2001 Jun 8;276(23):20108-15
references: 11274170
references: Oncogene. 2004 Jun 17;23(28):4873-84
references: 15064719
references: J Biol Chem. 2013 Mar 29;288(13):9438-46
references: 23396968
references: Genes Dev. 1995 May 1;9(9):1087-97
references: 7744250
references: Nature. 2002 Oct 17;419(6908):726-9
references: 12384700
references: Cancer Sci. 2008 Feb;99(2):202-8
references: 18271916
references: Mol Biol Cell. 2006 Aug;17(8):3345-55
references: 16707567
references: Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4044-51
references: 22343533