DIX Domains of Dvl and Axin Are Necessary for Protein Interactions and Their Ability To Regulate β-Catenin Stability

ABSTRACT The N-terminal region of Dvl-1 (a mammalian Dishevelled homolog) shares 37% identity with the C-terminal region of Axin, and this related region is named the DIX domain. The functions of the DIX domains of Dvl-1 and Axin were investigated. By yeast two-hybrid screening, the DIX domain of Dvl-1 was found to interact with Dvl-3, a second mammalian Dishevelled relative. The DIX domains of Dvl-1 and Dvl-3 directly bound one another. Furthermore, Dvl-1 formed a homo-oligomer. Axin also formed a homo-oligomer, and its DIX domain was necessary. The N-terminal region of Dvl-1, including its DIX domain, bound to Axin directly. Dvl-1 inhibited Axin-promoted glycogen synthase kinase 3β-dependent phosphorylation of β-catenin, and the DIX domain of Dvl-1 was required for this inhibitory activity. Expression of Dvl-1 in L cells induced the nuclear accumulation of β-catenin, and deletion of the DIX domain abolished this activity. Although expression of Axin in SW480 cells caused the degradation of β-catenin and reduced the cell growth rate, expression of an Axin mutant that lacks the DIX domain did not affect the level of β-catenin or the growth rate. These results indicate that the DIX domains of Dvl-1 and Axin are important for protein-protein interactions and that they are necessary for the ability of Dvl-1 and Axin to regulate the stability of β-catenin.

[1]  H. Varmus,et al.  Casein kinase 2 associates with and phosphorylates Dishevelled , 1997, The EMBO journal.

[2]  A. McMahon,et al.  Wnt genes and vertebrate development. , 1994, Current opinion in genetics & development.

[3]  T. Dale,et al.  Signal transduction by the Wnt family of ligands. , 1998, The Biochemical journal.

[4]  山本 英樹.  Axil, a member of the Axin family, interacts with both glycogen synthase kinase 3β and β-catenin and inhibits Axis formation of Xenopus embryos , 2001 .

[5]  Michael Kühl,et al.  Functional interaction of β-catenin with the transcription factor LEF-1 , 1996, Nature.

[6]  Akira Kikuchi,et al.  Axil, a Member of the Axin Family, Interacts with Both Glycogen Synthase Kinase 3β and β-Catenin and Inhibits Axis Formation ofXenopus Embryos , 1998, Molecular and Cellular Biology.

[7]  G. Struhl,et al.  Regulation of the Hedgehog and Wingless signalling pathways by the F-box/WD40-repeat protein Slimb , 1998, Nature.

[8]  Hans Clevers,et al.  XTcf-3 Transcription Factor Mediates β-Catenin-Induced Axis Formation in Xenopus Embryos , 1996, Cell.

[9]  M. Boutros,et al.  Dishevelled Activates JNK and Discriminates between JNK Pathways in Planar Polarity and wingless Signaling , 1998, Cell.

[10]  Akira Kikuchi,et al.  Axin, a negative regulator of the Wnt signaling pathway, forms a complex with GSK‐3β and β‐catenin and promotes GSK‐3β‐dependent phosphorylation of β‐catenin , 1998 .

[11]  A. Kikuchi,et al.  Tyrosine dephosphorylation of glycogen synthase kinase‐3 is involved in its extracellular signal‐dependent inactivation , 1996, FEBS letters.

[12]  J. Woodgett,et al.  Differential regulation of glycogen synthase kinase-3 beta by protein kinase C isotypes. , 1992, The Journal of biological chemistry.

[13]  J. Woodgett,et al.  Wingless inactivates glycogen synthase kinase‐3 via an intracellular signalling pathway which involves a protein kinase C. , 1996, The EMBO journal.

[14]  Douglas A. Melton,et al.  Injected Wnt RNA induces a complete body axis in Xenopus embryos , 1991, Cell.

[15]  T. Hinoi,et al.  Identification and Characterization of a Novel Protein Interacting with Ral-binding Protein 1, a Putative Effector Protein of Ral* , 1998, The Journal of Biological Chemistry.

[16]  Walter Birchmeier,et al.  Hot papers in cell biology - J. Behrens, J.P. von Kries, M. Kuehl, L. Bruhn, D. Wedlich, R. Grosschedl, W. Birchmeier: "Functional interaction of beta-catenin with the transcription factor LEF-1" - Comments by Walter Birchmeier , 1999 .

[17]  N. Perrimon,et al.  Conservation of dishevelled structure and function between flies and mice: isolation and characterization of Dvl2 , 1996, Mechanisms of Development.

[18]  S. Pierce,et al.  Regulation of Spemann organizer formation by the intracellular kinase Xgsk-3. , 1995, Development.

[19]  S. Gluecksohn‐Schoenheimer.  The effects of a lethal mutation responsible for duplications and twinning in mouse embryos. , 1949, The Journal of experimental zoology.

[20]  S. Sokol.  Analysis of Dishevelled signalling pathways during Xenopus development , 1996, Current Biology.

[21]  Hideki Yamamoto,et al.  Axin, a Negative Regulator of the Wnt Signaling Pathway, Directly Interacts with Adenomatous Polyposis Coli and Regulates the Stabilization of β-Catenin* , 1998, The Journal of Biological Chemistry.

[22]  L. Williams,et al.  Bridging of β-catenin and glycogen synthase kinase-3β by Axin and inhibition of β-catenin-mediated transcription , 1998 .

[23]  J. Klingensmith,et al.  The dishevelled protein is modified by wingless signaling in Drosophila. , 1995, Genes & development.

[24]  H. Theisen,et al.  dishevelled is required during wingless signaling to establish both cell polarity and cell identity. , 1994, Development.

[25]  R. Moon,et al.  The axis-inducing activity, stability, and subcellular distribution of beta-catenin is regulated in Xenopus embryos by glycogen synthase kinase 3. , 1996, Genes & development.

[26]  Wei Hsu,et al.  The Mouse Fused Locus Encodes Axin, an Inhibitor of the Wnt Signaling Pathway That Regulates Embryonic Axis Formation , 1997, Cell.

[27]  Jörg Stappert,et al.  β‐catenin is a target for the ubiquitin–proteasome pathway , 1997 .

[28]  Richard Paylor,et al.  Social Interaction and Sensorimotor Gating Abnormalities in Mice Lacking Dvl1 , 1997, Cell.

[29]  E. Krebs,et al.  Inactivation of Glycogen Synthase Kinase-3 by Epidermal Growth Factor Is Mediated by Mitogen-activated Protein Kinase/p90 Ribosomal Protein S6 Kinase Signaling Pathway in NIH/3T3 Cells (*) , 1995, The Journal of Biological Chemistry.

[30]  N. Perrimon,et al.  The Drosophila segment polarity gene dishevelled encodes a novel protein required for response to the wingless signal. , 1994, Genes & development.

[31]  Hans Clevers,et al.  Activation of β-Catenin-Tcf Signaling in Colon Cancer by Mutations in β-Catenin or APC , 1997, Science.

[32]  Paul Polakis,et al.  Downregulation of β-catenin by human Axin and its association with the APC tumor suppressor, β-catenin and GSK3β , 1998, Current Biology.

[33]  Jonathan A. Cooper,et al.  Mammalian Ras interacts directly with the serine/threonine kinase raf , 1993, Cell.

[34]  W. Birchmeier,et al.  Functional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta. , 1998, Science.

[35]  W. Nelson,et al.  Identification of Four Distinct Pools of Catenins in Mammalian Cells and Transformation-dependent Changes in Catenin Distributions among These Pools* , 1997, The Journal of Biological Chemistry.

[36]  N. Perrimon,et al.  Isolation and characterization of a mouse homolog of the Drosophila segment polarity gene dishevelled. , 1994, Developmental biology.

[37]  R. Nusse,et al.  Wnt signaling: a common theme in animal development. , 1997, Genes & development.

[38]  T. Uemura,et al.  Accumulation of Armadillo Induced by Wingless, Dishevelled, and Dominant-negative Zeste-white 3 Leads to Elevated DE-cadherin inDrosophila Clone 8 Wing Disc Cells* , 1997, The Journal of Biological Chemistry.

[39]  Paul Polakis,et al.  Binding of GSK3β to the APC-β-Catenin Complex and Regulation of Complex Assembly , 1996, Science.

[40]  T. Vasicek,et al.  Phenotypic and molecular analysis of a transgenic insertional allele of the mouse Fused locus. , 1995, Genetics.

[41]  S. Sokol,et al.  Axis determination in Xenopus involves biochemical interactions of axin, glycogen synthase kinase 3 and β-catenin , 1998, Current Biology.

[42]  R. Moon,et al.  Signal transduction through beta-catenin and specification of cell fate during embryogenesis. , 1996, Genes & development.

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

[44]  A. Kikuchi,et al.  Axin prevents Wnt-3a-induced accumulation of β-catenin , 1999, Oncogene.

[45]  Harold E. Varmus,et al.  Glycogen synthase kinase-3 and dorsoventral patterning in Xenopus embryos , 1995, Nature.

[46]  P. Polakis.  The adenomatous polyposis coli (APC) tumor suppressor. , 1997, Biochimica et biophysica acta.

[47]  N. Perrimon,et al.  Differential Recruitment of Dishevelled Provides Signaling Specificity in the Planar Cell Polarity and Wingless Signaling Pathways in Drosophila, Planar Cell Polarity (pcp) Signaling Is Mediated by the Receptor Frizzled (fz) and Transduced by Dishevelled (dsh). Wingless (wg) Signaling Also Requires , 2022 .

[48]  P. Cohen,et al.  Inactivation of glycogen synthase kinase-3 beta by phosphorylation: new kinase connections in insulin and growth-factor signalling. , 1993, The Biochemical journal.

[49]  K. Kinzler,et al.  Constitutive Transcriptional Activation by a β-Catenin-Tcf Complex in APC−/− Colon Carcinoma , 1997, Science.

[50]  B. Dallapiccola,et al.  cDNA characterization and chromosomal mapping of two human homologues of the Drosophila dishevelled polarity gene. , 1996, Human molecular genetics.

[51]  P. Cohen,et al.  Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B , 1995, Nature.