Wnt-5a/Ca2+-Induced NFAT Activity Is Counteracted by Wnt-5a/Yes-Cdc42-Casein Kinase 1α Signaling in Human Mammary Epithelial Cells

ABSTRACT Wnt-5a has been shown to influence the metastatic behavior of human breast cancer cells, and the loss of Wnt-5a expression is associated with metastatic disease. We show here that NFAT1, a transcription factor connected with breast cancer metastasis, is activated by Wnt-5a through a Ca2+ signaling pathway in human breast epithelial cells. This activation was simultaneously counteracted by a Wnt-5a-induced Yes/Cdc42 signaling pathway. The observation that inhibition of the Wnt-5a/Yes/Cdc42 signal prolonged the duration of ionomycin-induced NFAT1 activation revealed the general importance of this pathway. The Wnt-5a-induced inhibition of NFAT1 did not require glycogen synthase kinase 3β, JNK, or Pak1 activity or modulation of the cytoskeleton. Instead, we observed that Wnt-5a induced a complex formation of NFAT1/casein kinase 1α, even upon treatment with ionomycin, which was blocked upon inhibition of the Wnt-5a/Yes/Cdc42 signaling pathway. Our results explain why Wnt-5a/Ca2+-induced NFAT activity is hard to detect and suggest a novel mechanism by which Wnt-5a can suppress tumor-specific, agonist-induced NFAT activity and thus the metastatic behavior of breast cancer cells.

[1]  R. Nusse,et al.  Purified Wnt5a Protein Activates or Inhibits β-Catenin–TCF Signaling Depending on Receptor Context , 2006, PLoS biology.

[2]  A. Toker,et al.  Akt blocks breast cancer cell motility and invasion through the transcription factor NFAT. , 2005, Molecular cell.

[3]  T. Andersson,et al.  Wnt-5a protein expression in primary dukes B colon cancers identifies a subgroup of patients with good prognosis. , 2005, Cancer research.

[4]  Randall T. Moon,et al.  Wnt and calcium signaling: β-Catenin-independent pathways , 2005 .

[5]  H. Kondoh,et al.  Analysis of combinatorial effects of Wnts and Frizzleds on β‐catenin/armadillo stabilization and Dishevelled phosphorylation , 2005, Genes to cells : devoted to molecular & cellular mechanisms.

[6]  Andreas Gocht,et al.  The casein kinase 1 family: participation in multiple cellular processes in eukaryotes. , 2005, Cellular signalling.

[7]  G. Brabant,et al.  Wnt-5a has tumor suppressor activity in thyroid carcinoma , 2005, Oncogene.

[8]  O. Huber,et al.  A second protein kinase CK1-mediated step negatively regulates Wnt signalling by disrupting the lymphocyte enhancer factor-1/β-catenin complex , 2005, Cellular and Molecular Life Sciences.

[9]  P. Sobrado,et al.  Basic region of residues 228–231 of protein kinase CK1α is involved in its interaction with axin: Binding to axin does not affect the kinase activity , 2005, Journal of cellular biochemistry.

[10]  G. Landberg,et al.  Expression and signaling activity of Wnt-5a/discoidin domain receptor-1 and Syk plays distinct but decisive roles in breast cancer patient survival. , 2005, Clinical cancer research : an official journal of the American Association for Cancer Research.

[11]  Randall T Moon,et al.  Wnt and calcium signaling: beta-catenin-independent pathways. , 2005, Cell calcium.

[12]  Nam-Chul Ha,et al.  Mechanism of Phosphorylation-Dependent Binding of APC to β-Catenin and Its Role in β-Catenin Degradation , 2004 .

[13]  M. Okada,et al.  Activation of Cdc42 by trans interactions of the cell adhesion molecules nectins through c-Src and Cdc42-GEF FRG , 2004, The Journal of cell biology.

[14]  J. Qin,et al.  A Conserved Docking Motif for CK1 Binding Controls the Nuclear Localization of NFAT1 , 2004, Molecular and Cellular Biology.

[15]  U. Bommhardt,et al.  Active Protein Kinase B Regulates TCR Responsiveness by Modulating Cytoplasmic-Nuclear Localization of NFAT and NF-κB Proteins1 , 2004, The Journal of Immunology.

[16]  R. Nusse,et al.  Convergence of Wnt, ß-Catenin, and Cadherin Pathways , 2004, Science.

[17]  F. Rivas,et al.  Actin Cytoskeleton Regulates Calcium Dynamics and NFAT Nuclear Duration , 2004, Molecular and Cellular Biology.

[18]  R. Nusse,et al.  Convergence of Wnt, beta-catenin, and cadherin pathways. , 2004, Science.

[19]  Nam-Chul Ha,et al.  Mechanism of phosphorylation-dependent binding of APC to beta-catenin and its role in beta-catenin degradation. , 2004, Molecular cell.

[20]  T. Dale,et al.  Wnt Signaling and Mammary Tumorigenesis , 2004, Journal of Mammary Gland Biology and Neoplasia.

[21]  W. J. Wu,et al.  Epidermal Growth Factor-dependent Regulation of Cdc42 Is Mediated by the Src Tyrosine Kinase* , 2003, Journal of Biological Chemistry.

[22]  M. Nelson,et al.  Constitutively Elevated Nuclear Export Activity Opposes Ca2+-dependent NFATc3 Nuclear Accumulation in Vascular Smooth Muscle , 2003, Journal of Biological Chemistry.

[23]  C. Kuan,et al.  c‐Jun N‐terminal kinases (JNK) antagonize cardiac growth through cross‐talk with calcineurin–NFAT signaling , 2003, The EMBO journal.

[24]  Hosoon Choi,et al.  Wnt-5a inhibits the canonical Wnt pathway by promoting GSK-3–independent β-catenin degradation , 2003, The Journal of cell biology.

[25]  Jeffrey D. Axelrod,et al.  A Second Canon , 2003 .

[26]  Gilbert Weidinger,et al.  When Wnts antagonize Wnts , 2003, The Journal of cell biology.

[27]  Hans Clevers,et al.  Caught up in a Wnt storm: Wnt signaling in cancer. , 2003, Biochimica et biophysica acta.

[28]  R. Moon,et al.  Dishevelled activates Ca2+ flux, PKC, and CamKII in vertebrate embryos , 2003, The Journal of cell biology.

[29]  H. Varmus,et al.  Wnt/Wingless signaling through β-catenin requires the function of both LRP/Arrow and frizzled classes of receptors , 2003, BMC Cell Biology.

[30]  V. Braga,et al.  Tumor progression: Small GTPases and loss of cell-cell adhesion. , 2003, BioEssays : news and reviews in molecular, cellular and developmental biology.

[31]  J. Behrens,et al.  The Wnt signaling pathway and its role in tumor development , 2003, Journal of Cancer Research and Clinical Oncology.

[32]  A. Hall,et al.  Cdc42 regulates GSK-3β and adenomatous polyposis coli to control cell polarity , 2003, Nature.

[33]  T. Andersson,et al.  Wnt‐5a and G‐protein signaling are required for collagen‐induced DDR1 receptor activation and normal mammary cell adhesion , 2003, International journal of cancer.

[34]  Xi He,et al.  Coactivation of Rac and Rho by Wnt/Frizzled signaling is required for vertebrate gastrulation. , 2003, Genes & development.

[35]  Randall T Moon,et al.  A second canon. Functions and mechanisms of beta-catenin-independent Wnt signaling. , 2003, Developmental cell.

[36]  Minoti Hiremath,et al.  Beta-catenin and Tcfs in mammary development and cancer. , 2003, Journal of mammary gland biology and neoplasia.

[37]  M. Kühl,et al.  Increasingly complex: new players enter the Wnt signaling network. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[38]  Christopher C. W. Hughes,et al.  Endothelial Cells Stimulate T Cell NFAT Nuclear Translocation in the Presence of Cyclosporin A: Involvement of the wnt/Glycogen Synthase Kinase-3β Pathway1 , 2002, The Journal of Immunology.

[39]  Leslie M. Shaw,et al.  The role of NFAT transcription factors in integrin-mediated carcinoma invasion , 2002, Nature Cell Biology.

[40]  Antonio Vidal-Puig,et al.  Characterisation of the phosphorylation of beta-catenin at the GSK-3 priming site Ser45. , 2002, Biochemical and biophysical research communications.

[41]  Katsuhiko Mikoshiba,et al.  The Wnt/calcium pathway activates NF-AT and promotes ventral cell fate in Xenopus embryos , 2002, Nature.

[42]  G. Crabtree,et al.  NFAT Signaling Choreographing the Social Lives of Cells , 2002, Cell.

[43]  M. Bittner,et al.  Wnt5a signaling directly affects cell motility and invasion of metastatic melanoma. , 2002, Cancer cell.

[44]  C. Heldin,et al.  Transforming growth factor-beta-induced mobilization of actin cytoskeleton requires signaling by small GTPases Cdc42 and RhoA. , 2002, Molecular biology of the cell.

[45]  P. Bendahl,et al.  Loss of Wnt-5a protein is associated with early relapse in invasive ductal breast carcinomas. , 2002, Cancer research.

[46]  E. Nishida,et al.  JNK functions in the non‐canonical Wnt pathway to regulate convergent extension movements in vertebrates , 2002, EMBO reports.

[47]  Michael Kühl,et al.  Antagonistic regulation of convergent extension movements in Xenopus by Wnt/β-catenin and Wnt/Ca2+ signaling , 2001, Mechanisms of Development.

[48]  T. Andersson,et al.  Repression of Wnt-5a impairs DDR1 phosphorylation and modifies adhesion and migration of mammary cells. , 2001, Journal of cell science.

[49]  J Mao,et al.  Low-density lipoprotein receptor-related protein-5 binds to Axin and regulates the canonical Wnt signaling pathway. , 2001, Molecular cell.

[50]  R. Davis,et al.  Signal Transduction by the JNK Group of MAP Kinases , 2000, Cell.

[51]  J. Stam,et al.  Factor VIIa/Tissue Factor-induced Signaling via Activation of Src-like Kinases, Phosphatidylinositol 3-Kinase, and Rac* , 2000, The Journal of Biological Chemistry.

[52]  R. Moon,et al.  The Wnt/Ca2+ pathway: a new vertebrate Wnt signaling pathway takes shape. , 2000, Trends in genetics : TIG.

[53]  Juan Miguel Redondo,et al.  A Role for the p38 MAP Kinase Pathway in the Nuclear Shuttling of NFATp* , 2000, The Journal of Biological Chemistry.

[54]  E. Laue,et al.  Structure of Cdc42 bound to the GTPase binding domain of PAK , 2000, Nature Structural Biology.

[55]  R. Moon,et al.  Ca2+/Calmodulin-dependent Protein Kinase II Is Stimulated by Wnt and Frizzled Homologs and Promotes Ventral Cell Fates in Xenopus* , 2000, The Journal of Biological Chemistry.

[56]  M. Mandal,et al.  Regulation of Microfilament Reorganization and Invasiveness of Breast Cancer Cells by Kinase Dead p21-activated Kinase-1* , 2000, The Journal of Biological Chemistry.

[57]  N. Clipstone,et al.  Identification of Amino Acid Residues and Protein Kinases Involved in the Regulation of NFATc Subcellular Localization* , 2000, The Journal of Biological Chemistry.

[58]  J. Behrens,et al.  Biochemical interactions in the wnt pathway. , 2000, Biochimica et biophysica acta.

[59]  K. Matsumoto,et al.  Distinct Domains of Mouse Dishevelled Are Responsible for the c-Jun N-terminal Kinase/Stress-activated Protein Kinase Activation and the Axis Formation in Vertebrates* , 1999, The Journal of Biological Chemistry.

[60]  M. Yaffe,et al.  Affinity-driven peptide selection of an NFAT inhibitor more selective than cyclosporin A. , 1999, Science.

[61]  Jonathan A. Cooper,et al.  Src family kinases are required for integrin but not PDGFR signal transduction , 1999, The EMBO journal.

[62]  J. Loureiro,et al.  The Wnts , 1999, Current Biology.

[63]  J Mao,et al.  Dishevelled Proteins Lead to Two Signaling Pathways , 1999, The Journal of Biological Chemistry.

[64]  A. Weiss,et al.  A Nck‐Pak1 signaling module is required for T‐cell receptor‐mediated activation of NFAT, but not of JNK , 1998, The EMBO journal.

[65]  F. Shibasaki,et al.  Intramolecular Masking of Nuclear Import Signal on NF-AT4 by Casein Kinase I and MEKK1 , 1998, Cell.

[66]  D. Olson,et al.  Antisense wnt-5a mimics wnt-1-mediated C57MG mammary epithelial cell transformation. , 1998, Experimental cell research.

[67]  M Dickens,et al.  Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway. , 1997, Science.

[68]  K. Leach,et al.  Dynamic equilibrium between calcineurin and kinase activities regulates the phosphorylation state and localization of the nuclear factor of activated T-cells. , 1997, The Biochemical journal.

[69]  Christoph W. Turck,et al.  Nuclear Export of NF-ATc Enhanced by Glycogen Synthase Kinase-3 , 1997, Science.

[70]  J. Nathans,et al.  A Member of the Frizzled Protein Family Mediating Axis Induction by Wnt-5A , 1997, Science.

[71]  P. Hogan,et al.  Transcription factors of the NFAT family: regulation and function. , 1997, Annual review of immunology.

[72]  K. Leach,et al.  Direct Demonstration of NFATp Dephosphorylation and Nuclear Localization in Activated HT-2 Cells Using a Specific NFATp Polyclonal Antibody (*) , 1995, The Journal of Biological Chemistry.

[73]  M. Karin,et al.  Selective activation of the JNK signaling cascadeand c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs , 1995, Cell.

[74]  T. Hoey,et al.  Isolation of two new members of the NF-AT gene family and functional characterization of the NF-AT proteins. , 1995, Immunity.