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EMBO J. Sep 1, 1998; 17(17): 5129–5140.
PMCID: PMC1170841

Cloning and characterization of mCtBP2, a co-repressor that associates with basic Krüppel-like factor and other mammalian transcriptional regulators.

Abstract

Basic Krüppel-like factor (BKLF) is a zinc finger protein that recognizes CACCC elements in DNA. It is expressed highly in erythroid tissues, the brain and other selected cell types. We have studied the activity of BKLF and found that it is capable of repressing transcription, and have mapped its repression domain to the N-terminus. We carried out a two-hybrid screen against BKLF and isolated a novel clone encoding murine C-terminal-binding protein 2 (mCtBP2). mCtBP2 is related to human CtBP, a cellular protein which binds to a Pro-X-Asp-Leu-Ser motif in the C-terminus of the adenoviral oncoprotein, E1a. We show that mCtBP2 recognizes a related motif in the minimal repression domain of BKLF, and the integrity of this motif is required for repression activity. Moreover, when tethered to a promoter by a heterologous DNA-binding domain, mCtBP2 functions as a potent repressor. Finally, we demonstrate that mCtBP2 also interacts with the mammalian transcripition factors Evi-1, AREB6, ZEB and FOG. These results establish a new member of the CtBP family, mCtBP2, as a mammalian co-repressor targeting diverse transcriptional regulators.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson KP, Kern CB, Crable SC, Lingrel JB. Isolation of a gene encoding a functional zinc finger protein homologous to erythroid Krüppel-like factor: identification of a new multigene family. Mol Cell Biol. 1995 Nov;15(11):5957–5965. [PMC free article] [PubMed]
  • Bartholomew C, Kilbey A, Clark AM, Walker M. The Evi-1 proto-oncogene encodes a transcriptional repressor activity associated with transformation. Oncogene. 1997 Feb 6;14(5):569–577. [PubMed]
  • Boyd JM, Subramanian T, Schaeper U, La Regina M, Bayley S, Chinnadurai G. 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. 1993 Feb;12(2):469–478. [PMC free article] [PubMed]
  • Brehm A, Miska EA, McCance DJ, Reid JL, Bannister AJ, Kouzarides T. Retinoblastoma protein recruits histone deacetylase to repress transcription. Nature. 1998 Feb 5;391(6667):597–601. [PubMed]
  • Broihier HT, Moore LA, Van Doren M, Newman S, Lehmann R. zfh-1 is required for germ cell migration and gonadal mesoderm development in Drosophila. Development. 1998 Feb;125(4):655–666. [PubMed]
  • Chen C, Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PMC free article] [PubMed]
  • Choo Y, Sánchez-García I, Klug A. In vivo repression by a site-specific DNA-binding protein designed against an oncogenic sequence. Nature. 1994 Dec 15;372(6507):642–645. [PubMed]
  • Choo Y, Castellanos A, García-Hernández B, Sánchez-García I, Klug A. Promoter-specific activation of gene expression directed by bacteriophage-selected zinc fingers. J Mol Biol. 1997 Oct 31;273(3):525–532. [PubMed]
  • Crossley M, Whitelaw E, Perkins A, Williams G, Fujiwara Y, Orkin SH. Isolation and characterization of the cDNA encoding BKLF/TEF-2, a major CACCC-box-binding protein in erythroid cells and selected other cells. Mol Cell Biol. 1996 Apr;16(4):1695–1705. [PMC free article] [PubMed]
  • Fan CM, Maniatis T. Two different virus-inducible elements are required for human beta-interferon gene regulation. EMBO J. 1989 Jan;8(1):101–110. [PMC free article] [PubMed]
  • Feng WC, Southwood CM, Bieker JJ. Analyses of beta-thalassemia mutant DNA interactions with erythroid Krüppel-like factor (EKLF), an erythroid cell-specific transcription factor. J Biol Chem. 1994 Jan 14;269(2):1493–1500. [PubMed]
  • Fisher AL, Ohsako S, Caudy M. The WRPW motif of the hairy-related basic helix-loop-helix repressor proteins acts as a 4-amino-acid transcription repression and protein-protein interaction domain. Mol Cell Biol. 1996 Jun;16(6):2670–2677. [PMC free article] [PubMed]
  • Garrett-Sinha LA, Eberspaecher H, Seldin MF, de Crombrugghe B. A gene for a novel zinc-finger protein expressed in differentiated epithelial cells and transiently in certain mesenchymal cells. J Biol Chem. 1996 Dec 6;271(49):31384–31390. [PubMed]
  • Geisberg JV, Chen JL, Ricciardi RP. Subregions of the adenovirus E1A transactivation domain target multiple components of the TFIID complex. Mol Cell Biol. 1995 Nov;15(11):6283–6290. [PMC free article] [PubMed]
  • Genetta T, Ruezinsky D, Kadesch T. Displacement of an E-box-binding repressor by basic helix-loop-helix proteins: implications for B-cell specificity of the immunoglobulin heavy-chain enhancer. Mol Cell Biol. 1994 Sep;14(9):6153–6163. [PMC free article] [PubMed]
  • Goldberg JD, Yoshida T, Brick P. Crystal structure of a NAD-dependent D-glycerate dehydrogenase at 2.4 A resolution. J Mol Biol. 1994 Mar 4;236(4):1123–1140. [PubMed]
  • Haenlin M, Cubadda Y, Blondeau F, Heitzler P, Lutz Y, Simpson P, Ramain P. Transcriptional activity of pannier is regulated negatively by heterodimerization of the GATA DNA-binding domain with a cofactor encoded by the u-shaped gene of Drosophila. Genes Dev. 1997 Nov 15;11(22):3096–3108. [PMC free article] [PubMed]
  • Hanna-Rose W, Hansen U. Active repression mechanisms of eukaryotic transcription repressors. Trends Genet. 1996 Jun;12(6):229–234. [PubMed]
  • Heery DM, Kalkhoven E, Hoare S, Parker MG. A signature motif in transcriptional co-activators mediates binding to nuclear receptors. Nature. 1997 Jun 12;387(6634):733–736. [PubMed]
  • Hollenberg SM, Weinberger C, Ong ES, Cerelli G, Oro A, Lebo R, Thompson EB, Rosenfeld MG, Evans RM. Primary structure and expression of a functional human glucocorticoid receptor cDNA. Nature. 1985 Dec 19;318(6047):635–641. [PubMed]
  • Ikeda K, Halle JP, Stelzer G, Meisterernst M, Kawakami K. Involvement of negative cofactor NC2 in active repression by zinc finger-homeodomain transcription factor AREB6. Mol Cell Biol. 1998 Jan;18(1):10–18. [PMC free article] [PubMed]
  • Iwabuchi K, Li B, Bartel P, Fields S. Use of the two-hybrid system to identify the domain of p53 involved in oligomerization. Oncogene. 1993 Jun;8(6):1693–1696. [PubMed]
  • Katsanis N, Fisher EM. 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. 1998 Jan 15;47(2):294–299. [PubMed]
  • Kim JS, Pabo CO. Transcriptional repression by zinc finger peptides. Exploring the potential for applications in gene therapy. J Biol Chem. 1997 Nov 21;272(47):29795–29800. [PubMed]
  • Kim JS, Pabo CO. Getting a handhold on DNA: design of poly-zinc finger proteins with femtomolar dissociation constants. Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2812–2817. [PMC free article] [PubMed]
  • Kim JS, Kim J, Cepek KL, Sharp PA, Pabo CO. Design of TATA box-binding protein/zinc finger fusions for targeted regulation of gene expression. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3616–3620. [PMC free article] [PubMed]
  • Kuo CT, Veselits ML, Barton KP, Lu MM, Clendenin C, Leiden JM. The LKLF transcription factor is required for normal tunica media formation and blood vessel stabilization during murine embryogenesis. Genes Dev. 1997 Nov 15;11(22):2996–3006. [PMC free article] [PubMed]
  • Kuo CT, Veselits ML, Leiden JM. LKLF: A transcriptional regulator of single-positive T cell quiescence and survival. Science. 1997 Sep 26;277(5334):1986–1990. [PubMed]
  • Lee WS, Kao CC, Bryant GO, Liu X, Berk AJ. Adenovirus E1A activation domain binds the basic repeat in the TATA box transcription factor. Cell. 1991 Oct 18;67(2):365–376. [PubMed]
  • Lin RJ, Nagy L, Inoue S, Shao W, Miller WH, Jr, Evans RM. Role of the histone deacetylase complex in acute promyelocytic leukaemia. Nature. 1998 Feb 19;391(6669):811–814. [PubMed]
  • Magnaghi-Jaulin L, Groisman R, Naguibneva I, Robin P, Lorain S, Le Villain JP, Troalen F, Trouche D, Harel-Bellan A. Retinoblastoma protein represses transcription by recruiting a histone deacetylase. Nature. 1998 Feb 5;391(6667):601–605. [PubMed]
  • Merika M, Orkin SH. Functional synergy and physical interactions of the erythroid transcription factor GATA-1 with the Krüppel family proteins Sp1 and EKLF. Mol Cell Biol. 1995 May;15(5):2437–2447. [PMC free article] [PubMed]
  • Miller IJ, Bieker JJ. A novel, erythroid cell-specific murine transcription factor that binds to the CACCC element and is related to the Krüppel family of nuclear proteins. Mol Cell Biol. 1993 May;13(5):2776–2786. [PMC free article] [PubMed]
  • Mitchell PJ, Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. [PubMed]
  • Morishita K, Parker DS, Mucenski ML, Jenkins NA, Copeland NG, Ihle JN. Retroviral activation of a novel gene encoding a zinc finger protein in IL-3-dependent myeloid leukemia cell lines. Cell. 1988 Sep 9;54(6):831–840. [PubMed]
  • Morishita K, Suzukawa K, Taki T, Ihle JN, Yokota J. EVI-1 zinc finger protein works as a transcriptional activator via binding to a consensus sequence of GACAAGATAAGATAAN1-28 CTCATCTTC. Oncogene. 1995 May 18;10(10):1961–1967. [PubMed]
  • Mymryk JS. Tumour suppressive properties of the adenovirus 5 E1A oncogene. Oncogene. 1996 Oct 17;13(8):1581–1589. [PubMed]
  • Nagy L, Kao HY, Chakravarti D, Lin RJ, Hassig CA, Ayer DE, Schreiber SL, Evans RM. Nuclear receptor repression mediated by a complex containing SMRT, mSin3A, and histone deacetylase. Cell. 1997 May 2;89(3):373–380. [PubMed]
  • Nibu Y, Zhang H, Levine M. Interaction of short-range repressors with Drosophila CtBP in the embryo. Science. 1998 Apr 3;280(5360):101–104. [PubMed]
  • Nucifora G. The EVI1 gene in myeloid leukemia. Leukemia. 1997 Dec;11(12):2022–2031. [PubMed]
  • Nuez B, Michalovich D, Bygrave A, Ploemacher R, Grosveld F. Defective haematopoiesis in fetal liver resulting from inactivation of the EKLF gene. Nature. 1995 May 25;375(6529):316–318. [PubMed]
  • Perkins AC, Sharpe AH, Orkin SH. Lethal beta-thalassaemia in mice lacking the erythroid CACCC-transcription factor EKLF. Nature. 1995 May 25;375(6529):318–322. [PubMed]
  • Poortinga G, Watanabe M, Parkhurst SM. Drosophila CtBP: a Hairy-interacting protein required for embryonic segmentation and hairy-mediated transcriptional repression. EMBO J. 1998 Apr 1;17(7):2067–2078. [PMC free article] [PubMed]
  • Schaeper U, Boyd JM, Verma S, Uhlmann E, Subramanian T, Chinnadurai G. Molecular cloning and characterization of a cellular phosphoprotein that interacts with a conserved C-terminal domain of adenovirus E1A involved in negative modulation of oncogenic transformation. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10467–10471. [PMC free article] [PubMed]
  • Schüle R, Muller M, Otsuka-Murakami H, Renkawitz R. Cooperativity of the glucocorticoid receptor and the CACCC-box binding factor. Nature. 1988 Mar 3;332(6159):87–90. [PubMed]
  • Shields JM, Christy RJ, Yang VW. Identification and characterization of a gene encoding a gut-enriched Krüppel-like factor expressed during growth arrest. J Biol Chem. 1996 Aug 16;271(33):20009–20017. [PMC free article] [PubMed]
  • Sollerbrant K, Chinnadurai G, Svensson C. The CtBP binding domain in the adenovirus E1A protein controls CR1-dependent transactivation. Nucleic Acids Res. 1996 Jul 1;24(13):2578–2584. [PMC free article] [PubMed]
  • Strähle U, Schmid W, Schütz G. Synergistic action of the glucocorticoid receptor with transcription factors. EMBO J. 1988 Nov;7(11):3389–3395. [PMC free article] [PubMed]
  • Taguchi H, Ohta T. Histidine 296 is essential for the catalysis in Lactobacillus plantarum D-lactate dehydrogenase. J Biol Chem. 1993 Aug 25;268(24):18030–18034. [PubMed]
  • Tanaka T, Mitani K, Kurokawa M, Ogawa S, Tanaka K, Nishida J, Yazaki Y, Shibata Y, Hirai H. Dual functions of the AML1/Evi-1 chimeric protein in the mechanism of leukemogenesis in t(3;21) leukemias. Mol Cell Biol. 1995 May;15(5):2383–2392. [PMC free article] [PubMed]
  • Tsang AP, Visvader JE, Turner CA, Fujiwara Y, Yu C, Weiss MJ, Crossley M, Orkin SH. FOG, a multitype zinc finger protein, acts as a cofactor for transcription factor GATA-1 in erythroid and megakaryocytic differentiation. Cell. 1997 Jul 11;90(1):109–119. [PubMed]
  • Watanabe Y, Kawakami K, Hirayama Y, Nagano K. Transcription factors positively and negatively regulating the Na,K-ATPase alpha 1 subunit gene. J Biochem. 1993 Dec;114(6):849–855. [PubMed]
  • Yang XJ, Ogryzko VV, Nishikawa J, Howard BH, Nakatani Y. A p300/CBP-associated factor that competes with the adenoviral oncoprotein E1A. Nature. 1996 Jul 25;382(6589):319–324. [PubMed]
  • Yet SF, McA'Nulty MM, Folta SC, Yen HW, Yoshizumi M, Hsieh CM, Layne MD, Chin MT, Wang H, Perrella MA, et al. Human EZF, a Krüppel-like zinc finger protein, is expressed in vascular endothelial cells and contains transcriptional activation and repression domains. J Biol Chem. 1998 Jan 9;273(2):1026–1031. [PubMed]
  • Zamanian M, La Thangue NB. Adenovirus E1a prevents the retinoblastoma gene product from repressing the activity of a cellular transcription factor. EMBO J. 1992 Jul;11(7):2603–2610. [PMC free article] [PubMed]

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