Members of the Tcf/Lef family of the HMG box transcription factors are nuclear effectors of the Wnt signal transduction pathway. Upon Wnt signaling, TCF/LEF proteins interact with beta-catenin and activate transcription of target genes, while, in the absence of the Wnt signal, TCFs function as transcriptional repressors. All vertebrate Tcf/Lef transcription factors associate with TLE/Groucho-related co-repressors, and here we provide evidence for an interaction between the C-terminus of the TCF-4 HMG box protein and the C-terminal binding protein 1 (CtBP1) transcriptional co-repressor. Using Wnt-1-stimulated human embryonic kidney 293 cells, we show that CtBP1 represses the transcriptional activity of a Tcf/beta-catenin-dependent synthetic promoter and, furthermore, decreases the expression of the endogenous Wnt target, Axin2/Conductin. The CtBP1-mediated repression was alleviated by trichostatin A treatment, indicating that the CtBP inhibitory mechanism is dependent on the activity of histone deacetylases.

Institute of Molecular Genetics Academy of Sciences of the Czech Republic Videnska 1083 142 20 Prague 4 Czech Republic

Zobrazit více v PubMed

Cadigan K.M. and Nusse,R. (1997) Wnt signaling: a common theme in animal development. Genes Dev., 11, 3286–3305. PubMed

Bienz M. and Clevers,H. (2000) Linking colorectal cancer to Wnt signaling. Cell, 103, 311–320. PubMed

Miller J.R. and Moon,R.T. (1996) Signal transduction through beta-catenin and specification of cell fate during embryogenesis. Genes Dev., 10, 2527–2539. PubMed

Coates J.C., Grimson,M.J., Williams,R.S., Bergman,W., Blanton,R.L. and Harwood,A.J. (2002) Loss of the beta-catenin homologue aardvark causes ectopic stalk formation in Dictyostelium. Mech. Dev., 116, 117–127. PubMed

Hobmayer B., Rentzsch,F., Kuhn,K., Happel,C.M., von Laue,C.C., Snyder,P., Rothbacher,U. and Holstein,T.W. (2000) WNT signalling molecules act in axis formation in the diploblastic metazoan Hydra. Nature, 407, 186–189. PubMed

Grimson M.J., Coates,J.C., Reynolds,J.P., Shipman,M., Blanton,R.L. and Harwood,A.J. (2000) Adherens junctions and beta-catenin-mediated cell signalling in a non-metazoan organism. Nature, 408, 727–731. PubMed

Pinson K.I., Brennan,J., Monkley,S., Avery,B.J. and Skarnes,W.C. (2000) An LDL-receptor-related protein mediates Wnt signalling in mice. Nature, 407, 535–538. PubMed

Tamai K., Semenov,M., Kato,Y., Spokony,R., Liu,C., Katsuyama,Y., Hess,F., Saint-Jeannet,J.P. and He,X. (2000) LDL-receptor-related proteins in Wnt signal transduction. Nature, 407, 530–535. PubMed

Wehrli M., Dougan,S.T., Caldwell,K., O’Keefe,L., Schwartz,S., Vaizel-Ohayon,D., Schejter,E., Tomlinson,A. and DiNardo,S. (2000) arrow encodes an LDL-receptor-related protein essential for Wingless signalling. Nature, 407, 527–530. PubMed

Aberle H., Bauer,A., Stappert,J., Kispert,A. and Kemler,R. (1997) beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J., 16, 3797–3804. PubMed PMC

Jiang J. and Struhl,G. (1998) Regulation of the Hedgehog and Wingless signalling pathways by the F-box/WD40-repeat protein Slimb. Nature, 391, 493–496. PubMed

Marikawa Y. and Elinson,R.P. (1998) beta-TrCP is a negative regulator of Wnt/beta-catenin signaling pathway and dorsal axis formation in Xenopus embryos. Mech. Dev., 77, 75–80. PubMed

Hart M., Concordet,J.P., Lassot,I., Albert,I., del los Santos,R., Durand,H., Perret,C., Rubinfeld,B., Margottin,F., Benarous,R. et al. (1999) The F-box protein beta-TrCP associates with phosphorylated beta-catenin and regulates its activity in the cell. Curr. Biol., 9, 207–210. PubMed

Hart M.J., de los Santos,R., Albert,I.N., Rubinfeld,B. and Polakis,P. (1998) Downregulation of beta-catenin by human Axin and its association with the APC tumor suppressor, beta-catenin and GSK3 beta. Curr. Biol., 8, 573–581. PubMed

Fagotto F., Jho,E., Zeng,L., Kurth,T., Joos,T., Kaufmann,C. and Costantini,F. (1999) Domains of axin involved in protein–protein interactions, Wnt pathway inhibition, and intracellular localization. J. Cell Biol., 145, 741–756. PubMed PMC

Liu C., Li,Y., Semenov,M., Han,C., Baeg,G.H., Tan,Y., Zhang,Z., Lin,X. and He,X. (2002) Control of beta-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell, 108, 837–847. PubMed

Yanagawa S., Matsuda,Y., Lee,J.S., Matsubayashi,H., Sese,S., Kadowaki,T. and Ishimoto,A. (2002) Casein kinase I phosphorylates the Armadillo protein and induces its degradation in Drosophila. EMBO J., 21, 1733–1742. PubMed PMC

Behrens J., Jerchow,B.A., Wurtele,M., Grimm,J., Asbrand,C., Wirtz,R., Kuhl,M., Wedlich,D. and Birchmeier,W. (1998) Functional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta. Science, 280, 596–599. PubMed

Kishida M., Koyama,S., Kishida,S., Matsubara,K., Nakashima,S., Higano,K., Takada,R., Takada,S. and Kikuchi,A. (1999) Axin prevents Wnt-3a-induced accumulation of beta-catenin. Oncogene, 18, 979–985. PubMed

Zeng L., Fagotto,F., Zhang,T., Hsu,W., Vasicek,T.J., Perry,W.L.,3rd, Lee,J.J., Tilghman,S.M., Gumbiner,B.M. and Costantini,F. (1997) The mouse Fused locus encodes Axin, an inhibitor of the Wnt signaling pathway that regulates embryonic axis formation. Cell, 90, 181–192. PubMed

Boutros M., Mihaly,J., Bouwmeester,T. and Mlodzik,M. (2000) Signaling specificity by Frizzled receptors in Drosophila. Science, 288, 1825–1828. PubMed

Axelrod J.D., Miller,J.R., Shulman,J.M., Moon,R.T. and Perrimon,N. (1998) Differential recruitment of Dishevelled provides signaling specificity in the planar cell polarity and Wingless signaling pathways. Genes Dev., 12, 2610–2622. PubMed PMC

Huber O., Korn,R., McLaughlin,J., Ohsugi,M., Herrmann,B.G. and Kemler,R. (1996) Nuclear localization of beta-catenin by interaction with transcription factor LEF-1. Mech. Dev., 59, 3–10. PubMed

Behrens J., von Kries,J.P., Kuhl,M., Bruhn,L., Wedlich,D., Grosschedl,R. and Birchmeier,W. (1996) Functional interaction of beta-catenin with the transcription factor LEF-1. Nature, 382, 638–642. PubMed

Molenaar M., Roose,J., Peterson,J., Venanzi,S., Clevers,H. and Destree,O. (1998) Differential expression of the HMG box transcription factors XTcf-3 and XLef-1 during early Xenopus development. Mech. Dev., 75, 151–154. PubMed

Morin P.J., Sparks,A.B., Korinek,V., Barker,N., Clevers,H., Vogelstein,B. and Kinzler,K.W. (1997) Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science, 275, 1787–1790. PubMed

Korinek V., Barker,N., Morin,P.J., van Wichen,D., de Weger,R., Kinzler,K.W., Vogelstein,B. and Clevers,H. (1997) Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC–/– colon carcinoma. Science, 275, 1784–1787. PubMed

He T.C., Sparks,A.B., Rago,C., Hermeking,H., Zawel,L., da Costa,L.T., Morin,P.J., Vogelstein,B. and Kinzler,K.W. (1998) Identification of c-MYC as a target of the APC pathway. Science, 281, 1509–1512. PubMed

Kioussi C., Briata,P., Baek,S.H., Rose,D.W., Hamblet,N.S., Herman,T., Ohgi,K.A., Lin,C., Gleiberman,A., Wang,J. et al. (2002) Identification of a Wnt/Dvl/beta-catenin → Pitx2 pathway mediating cell-type-specific proliferation during development. Cell, 111, 673–685. PubMed

Shtutman M., Zhurinsky,J., Simcha,I., Albanese,C., D’Amico,M., Pestell,R. and Ben-Ze’ev,A. (1999) The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc. Natl Acad. Sci. USA, 96, 5522–5527. PubMed PMC

Tetsu O. and McCormick,F. (1999) Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature, 398, 422–426. PubMed

Fedi P., Bafico,A., Nieto Soria,A., Burgess,W.H., Miki,T., Bottaro,D.P., Kraus,M.H. and Aaronson,S.A. (1999) Isolation and biochemical characterization of the human Dkk-1 homologue, a novel inhibitor of mammalian Wnt signaling. J. Biol. Chem., 274, 19465–19472. PubMed

Glinka A., Wu,W., Delius,H., Monaghan,A.P., Blumenstock,C. and Niehrs,C. (1998) Dickkopf-1 is a member of a new family of secreted proteins and functions in head induction. Nature, 391, 357–362. PubMed

Hsieh J.C., Kodjabachian,L., Rebbert,M.L., Rattner,A., Smallwood,P.M., Samos,C.H., Nusse,R., Dawid,I.B. and Nathans,J. (1999) A new secreted protein that binds to Wnt proteins and inhibits their activities. Nature, 398, 431–436. PubMed

Rattner A., Hsieh,J.C., Smallwood,P.M., Gilbert,D.J., Copeland,N.G., Jenkins,N.A. and Nathans,J. (1997) A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors. Proc. Natl Acad. Sci. USA, 94, 2859–2863. PubMed PMC

Leyns L., Bouwmeester,T., Kim,S.H., Piccolo,S. and De Robertis,E.M. (1997) Frzb-1 is a secreted antagonist of Wnt signaling expressed in the Spemann organizer. Cell, 88, 747–756. PubMed PMC

Melkonyan H.S., Chang,W.C., Shapiro,J.P., Mahadevappa,M., Fitzpatrick,P.A., Kiefer,M.C., Tomei,L.D. and Umansky,S.R. (1997) SARPs: a family of secreted apoptosis-related proteins. Proc. Natl Acad. Sci. USA, 94, 13636–13641. PubMed PMC

Rocheleau C.E., Yasuda,J., Shin,T.H., Lin,R., Sawa,H., Okano,H., Priess,J.R., Davis,R.J. and Mello,C.C. (1999) WRM-1 activates the LIT-1 protein kinase to transduce anterior/posterior polarity signals in C.elegans. Cell, 97, 717–726. PubMed

Ishitani T., Ninomiya-Tsuji,J., Nagai,S., Nishita,M., Meneghini,M., Barker,N., Waterman,M., Bowerman,B., Clevers,H., Shibuya,H. et al. (1999) The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF. Nature, 399, 798–802. PubMed

Meneghini M.D., Ishitani,T., Carter,J.C., Hisamoto,N., Ninomiya-Tsuji,J., Thorpe,C.J., Hamill,D.R., Matsumoto,K. and Bowerman,B. (1999) MAP kinase and Wnt pathways converge to downregulate an HMG-domain repressor in Caenorhabditis elegans. Nature, 399, 793–797. PubMed

Bauer A., Chauvet,S., Huber,O., Usseglio,F., Rothbacher,U., Aragnol,D., Kemler,R. and Pradel,J. (2000) Pontin52 and reptin52 function as antagonistic regulators of beta-catenin signalling activity. EMBO J., 19, 6121–6130. PubMed PMC

Barker N., Hurlstone,A., Musisi,H., Miles,A., Bienz,M. and Clevers,H. (2001) The chromatin remodelling factor Brg-1 interacts with beta-catenin to promote target gene activation. EMBO J., 20, 4935–4943. PubMed PMC

Hecht A., Vleminckx,K., Stemmler,M.P., van Roy,F. and Kemler,R. (2000) The p300/CBP acetyltransferases function as transcriptional coactivators of beta-catenin in vertebrates. EMBO J., 19, 1839–1850. PubMed PMC

Kramps T., Peter,O., Brunner,E., Nellen,D., Froesch,B., Chatterjee,S., Murone,M., Zullig,S. and Basler,K. (2002) Wnt/wingless signaling requires BCL9/legless-mediated recruitment of pygopus to the nuclear beta-catenin-TCF complex. Cell, 109, 47–60. PubMed

Parker D.S., Jemison,J. and Cadigan,K.M. (2002) Pygopus, a nuclear PHD-finger protein required for Wingless signaling in Drosophila. Development, 129, 2565–2576. PubMed

Zorn A.M., Barish,G.D., Williams,B.O., Lavender,P., Klymkowsky,M.W. and Varmus,H.E. (1999) Regulation of Wnt signaling by Sox proteins: XSox17 alpha/beta and XSox3 physically interact with beta-catenin. Mol. Cell, 4, 487–498. PubMed

Tago K., Nakamura,T., Nishita,M., Hyodo,J., Nagai,S., Murata,Y., Adachi,S., Ohwada,S., Morishita,Y., Shibuya,H. et al. (2000) Inhibition of Wnt signaling by ICAT, a novel beta-catenin-interacting protein. Genes Dev., 14, 1741–1749. PubMed PMC

Clevers H. and van de Wetering,M. (1997) TCF/LEF factor earn their wings. Trends Genet., 13, 485–489. PubMed

Eastman Q. and Grosschedl,R. (1999) Regulation of LEF-1/TCF transcription factors by Wnt and other signals. Curr. Opin. Cell Biol., 11, 233–240. PubMed

Roose J., Molenaar,M., Peterson,J., Hurenkamp,J., Brantjes,H., Moerer,P., van de Wetering,M., Destree,O. and Clevers,H. (1998) The Xenopus Wnt effector XTcf-3 interacts with Groucho-related transcriptional repressors. Nature, 395, 608–612. PubMed

Chen G., Fernandez,J., Mische,S. and Courey,A.J. (1999) A functional interaction between the histone deacetylase Rpd3 and the corepressor groucho in Drosophila development. Genes Dev., 13, 2218–2230. PubMed PMC

Brantjes H., Roose,J., van De Wetering,M. and Clevers,H. (2001) All Tcf HMG box transcription factors interact with Groucho-related co-repressors. Nucleic Acids Res., 29, 1410–1419. PubMed PMC

van Genderen C., Okamura,R.M., Farinas,I., Quo,R.G., Parslow,T.G., Bruhn,L. and Grosschedl,R. (1994) Development of several organs that require inductive epithelial-mesenchymal interactions is impaired in LEF-1-deficient mice. Genes Dev., 8, 2691–2703. PubMed

Verbeek S., Izon,D., Hofhuis,F., Robanus-Maandag,E., te Riele,H., van de Wetering,M., Oosterwegel,M., Wilson,A., MacDonald,H.R. and Clevers,H. (1995) An HMG-box-containing T-cell factor required for thymocyte differentiation. Nature, 374, 70–74. PubMed

Korinek V., Barker,N., Moerer,P., van Donselaar,E., Huls,G., Peters,P.J. and Clevers,H. (1998) Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4. Nature Genet., 19, 379–383. PubMed

Yasumoto K., Takeda,K., Saito,H., Watanabe,K., Takahashi,K. and Shibahara,S. (2002) Microphthalmia-associated transcription factor interacts with LEF-1, a mediator of Wnt signaling. EMBO J., 21, 2703–2714. PubMed PMC

Bruhn L., Munnerlyn,A. and Grosschedl,R. (1997) ALY, a context-dependent coactivator of LEF-1 and AML-1, is required for TCRalpha enhancer function. Genes Dev., 11, 640–653. PubMed

Sachdev S., Bruhn,L., Sieber,H., Pichler,A., Melchior,F. and Grosschedl,R. (2001) PIASy, a nuclear matrix-associated SUMO E3 ligase, represses LEF1 activity by sequestration into nuclear bodies. Genes Dev., 15, 3088–3103. PubMed PMC

Billin A.N., Thirlwell,H. and Ayer,D.E. (2000) Beta-catenin-histone deacetylase interactions regulate the transition of LEF1 from a transcriptional repressor to an activator. Mol. Cell. Biol., 20, 6882–6890. PubMed PMC

Lee E., Salic,A. and Kirschner,M.W. (2001) Physiological regulation of [beta]-catenin stability by Tcf3 and CK1epsilon. J. Cell Biol., 154, 983–993. PubMed PMC

Brannon M., Brown,J.D., Bates,R., Kimelman,D. and Moon,R.T. (1999) XCtBP is a XTcf-3 co-repressor with roles throughout Xenopus development. Development, 126, 3159–3170. PubMed

Hecht A. and Stemmler,M.P. (2002) Identification of a promoter-specific transcriptional activation domain at the C-terminus of the Wnt-effector protein TCF4. J. Biol. Chem., 22, 22. PubMed

Hovanes K., Li,T.W., Munguia,J.E., Truong,T., Milovanovic,T., Lawrence Marsh,J., Holcombe,R.F. and Waterman,M.L. (2001) Beta-catenin-sensitive isoforms of lymphoid enhancer factor-1 are selectively expressed in colon cancer. Nature Genet., 28, 53–57. PubMed

Roose J., Huls,G., van Beest,M., Moerer,P., van der Horn,K., Goldschmeding,R., Logtenberg,T. and Clevers,H. (1999) Synergy between tumor suppressor APC and the beta-catenin-Tcf4 target Tcf1. Science, 285, 1923–1926. PubMed

Gradl D., Konig,A. and Wedlich,D. (2002) Functional diversity of Xenopus lymphoid enhancer factor/T-cell factor transcription factors relies on combinations of activating and repressing elements. J. Biol. Chem., 277, 14159–14171. PubMed

Hoshimaru M., Ray,J., Sah,D.W. and Gage,F.H. (1996) Differentiation of the immortalized adult neuronal progenitor cell line HC2S2 into neurons by regulatable suppression of the v-myc oncogene. Proc. Natl Acad. Sci. USA, 93, 1518–1523. PubMed PMC

Dignam J.D., Lebovitz,R.M. and Roeder,R.G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res., 11, 1475–1489. PubMed PMC

Chomczynski P. and Sacchi,N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem., 162, 156–159. PubMed

Vandesompele J., De Preter,K., Pattyn,F., Poppe,B., Van Roy,N., De Paepe,A. and Speleman,F. (2002) Accurate normalization of real-time quantitative RT–PCR data by geometric averaging of multiple internal control genes. Genome Biol., 3, RESEARCH0034. PubMed PMC

Boyd J.M., Subramanian,T., Schaeper,U., La Regina,M., Bayley,S. and Chinnadurai,G. (1993) A region in the C-terminus of adenovirus 2/5 E1a protein is required for association with a cellular phosphoprotein and important for the negative modulation of T24-ras mediated transformation, tumorigenesis and metastasis. EMBO J., 12, 469–478. PubMed PMC

Katsanis N. and Fisher,E.M. (1998) A novel C-terminal binding protein (CTBP2) is closely related to CTBP1, an adenovirus E1A-binding protein, and maps to human chromosome 21q21.3. Genomics, 47, 294–299. PubMed

Chinnadurai G. (2002) CtBP, an unconventional transcriptional corepressor in development and oncogenesis. Mol. Cell, 9, 213–224. PubMed

Korinek V., Barker,N., Willert,K., Molenaar,M., Roose,J., Wagenaar,G., Markman,M., Lamers,W., Destree,O. and Clevers,H. (1998) Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. Mol. Cell. Biol., 18, 1248–1256. PubMed PMC

Van de Wetering M., Castrop,J., Korinek,V. and Clevers,H. (1996) Extensive alternative splicing and dual promoter usage generate Tcf-1 protein isoforms with differential transcription control properties. Mol. Cell. Biol., 16, 745–752. PubMed PMC

Koipally J. and Georgopoulos,K. (2000) Ikaros interactions with CtBP reveal a repression mechanism that is independent of histone deacetylase activity. J. Biol. Chem., 275, 19594–19602. PubMed

Leung J.Y., Kolligs,F.T., Wu,R., Zhai,Y., Kuick,R., Hanash,S., Cho,K.R. and Fearon,E.R. (2002) Activation of AXIN2 expression by beta-catenin-T cell factor. A feedback repressor pathway regulating Wnt signaling. J. Biol. Chem., 277, 21657–21665. PubMed

Lustig B., Jerchow,B., Sachs,M., Weiler,S., Pietsch,T., Karsten,U., van de Wetering,M., Clevers,H., Schlag,P.M., Birchmeier,W. et al. (2002) Negative feedback loop of Wnt signaling through upregulation of conductin/axin2 in colorectal and liver tumors. Mol. Cell. Biol., 22, 1184–1193. PubMed PMC

Hildebrand J.D. and Soriano,P. (2002) Overlapping and unique roles for C-terminal binding protein 1 (CtBP1) and CtBP2 during mouse development. Mol. Cell. Biol., 22, 5296–5307. PubMed PMC

Porfiri E., Rubinfeld,B., Albert,I., Hovanes,K., Waterman,M. and Polakis,P. (1997) Induction of a beta-catenin-LEF-1 complex by wnt-1 and transforming mutants of beta-catenin. Oncogene, 15, 2833–2839. PubMed

Pukrop T., Gradl,D., Henningfeld,K.A., Knochel,W., Wedlich,D. and Kuhl,M. (2001) Identification of two regulatory elements within the high mobility group box transcription factor XTCF-4. J. Biol. Chem., 276, 8968–8978. PubMed

Zhang H., Levine,M. and Ashe,H.L. (2001) Brinker is a sequence-specific transcriptional repressor in the Drosophila embryo. Genes Dev., 15, 261–266. PubMed PMC

Poortinga G., Watanabe,M. and Parkhurst,S.M. (1998) Drosophila CtBP: a Hairy-interacting protein required for embryonic segmentation and hairy-mediated transcriptional repression. EMBO J., 17, 2067–2078. PubMed PMC

Sundqvist A., Sollerbrant,K. and Svensson,C. (1998) The carboxy-terminal region of adenovirus E1A activates transcription through targeting of a C-terminal binding protein-histone deacetylase complex. FEBS Lett., 429, 183–188. PubMed

Kumar V., Carlson,J.E., Ohgi,K.A., Edwards,T.A., Rose,D.W., Escalante,C.R., Rosenfeld,M.G. and Aggarwal,A.K. (2002) Transcription corepressor CtBP is an NAD(+)-regulated dehydrogenase. Mol. Cell, 10, 857–869. PubMed

Zhang Q., Piston,D.W. and Goodman,R.H. (2002) Regulation of corepressor function by nuclear NADH. Science, 295, 1895–1897. PubMed

Multifaceted roles for BCL3 in cancer: a proto-oncogene comes of age

Wnt Effector TCF4 Is Dispensable for Wnt Signaling in Human Cancer Cells

TCF/LEF Transcription Factors: An Update from the Internet Resources

Genetic Evidence for Function of the bHLH-PAS Protein Gce/Met As a Juvenile Hormone Receptor

Dazap2 modulates transcription driven by the Wnt effector TCF-4

HIC1 attenuates Wnt signaling by recruitment of TCF-4 and beta-catenin to the nuclear bodies