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Biochemical characterization of the zinc-finger protein 217 transcriptional repressor complex: identification of a ZNF217 consensus recognition sequence

Oncogene volume 26, pages 3378–3386 (2007)Cite this article

Abstract

Zinc-finger protein 217 (ZNF217) is a Kruppel-like zinc-finger protein located at 20q13.2, within a region of recurrent maximal amplification. Here, we demonstrate that ZNF217 is a transcriptional repressor protein and report the purification and characterization of a ZNF217 complex. The purified ZNF217 complex consists of approximately six proteins and contains the transcriptional co-repressors CoREST, BHC110/LSD1, histone deacetylase (HDAC) 2 and C-terminal binding protein (CtBP1). The purified ZNF217 complex possesses deacetylase activity as well as lysine 4 histone H3-specific demethylase activity that is most likely mediated by the BHC110/LSD1 component. To determine if ZNF217 is a sequence-specific binding protein, we have made use of cyclic amplification and selection of targets (CAST) assay and identify for the first time a ZNF217 DNA consensus recognition sequence (CRS) that is highly conserved in the human E-cadherin promoter. Chromatin immunoprecipitation (ChIP) experiments demonstrate that ZNF217, as well as the other components of the ZNF217 complex, are found on the region of the proximal E-cadherin promoter that contains the identified ZNF217 CRS in vivo. Using a combination of transient transfections and small interfering RNA, we demonstrate that ZNF217 represses the E-cadherin promoter. Collectively, our results implicate ZNF217 and its associated proteins in a novel pathway that may have profound effects on cancer progression.

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References

  • Andres ME, Burger C, Peral-Rubio MJ, Battaglioli E, Anderson ME, Grimes J et al. (1999). CoREST: a functional corepressor required for regulation of neural-specific gene expression. Proc Natl Acad Sci USA 96: 9873–9878.

    Article  CAS  Google Scholar 

  • Ausubel FM . (2002). Current Protocols in Molecular Biology. John Wiley: New York.

    Google Scholar 

  • Batlle E, Sancho E, Franci C, Dominguez D, Monfar M, Baulida J et al. (2000). The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol 2: 84–89.

    Article  CAS  Google Scholar 

  • Bukholm IK, Nesland JM, Borresen-Dale AL . (2000). Re-expression of E-cadherin, alpha-catenin and beta-catenin, but not of gamma-catenin, in metastatic tissue from breast cancer patients. J Pathol 190: 15–19.

    Article  CAS  Google Scholar 

  • Cano A, Perez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG et al. (2000). The transcription factor snail controls epithelial–mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2: 76–83.

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Collins C, Rommens JM, Kowbel D, Godfrey T, Tanner M, Hwang SI et al. (1998). Positional cloning of ZNF217 and NABC1: genes amplified at 20q13.2 and overexpressed in breast carcinoma. Proc Natl Acad Sci USA 95: 8703–8708.

    Article  CAS  Google Scholar 

  • Collins C, Volik S, Kowbel D, Ginzinger D, Ylstra B, Cloutier T et al. (2001). Comprehensive genome sequence analysis of a breast cancer amplicon. Genome Res 11: 1034–1042.

    Article  CAS  Google Scholar 

  • Comijn J, Berx G, Vermassen P, Verschueren K, van Grunsven L, Bruyneel E et al. (2001). The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. Mol Cell 7: 1267–1278.

    Article  CAS  Google Scholar 

  • Dellaire G, Makarov EM, Cowger JJ, Longman D, Sutherland HG, Luhrmann R et al. (2002). Mammalian PRP4 kinase copurifies and interacts with components of both the U5 snRNP and the N-CoR deacetylase complexes. Mol Cell Biol 22: 5141–5156.

    Article  CAS  Google Scholar 

  • Elrod-Erickson M, Pabo CO . (1999). Binding studies with mutants of Zif268. Contribution of individual side chains to binding affinity and specificity in the Zif268 zinc finger–DNA complex. J Biol Chem 274: 19281–19285.

    Article  CAS  Google Scholar 

  • Grimes JA, Nielsen SJ, Battaglioli E, Miska EA, Speh JC, Berry DL et al. (2000). The co-repressor mSin3A is a functional component of the REST–CoREST repressor complex. J Biol Chem 275: 9461–9467.

    Article  CAS  Google Scholar 

  • Grooteclaes ML, Frisch SM . (2000). Evidence for a function of CtBP in epithelial gene regulation and anoikis. Oncogene 19: 3823–3828.

    Article  CAS  Google Scholar 

  • Hakimi MA, Dong Y, Lane WS, Speicher DW, Shiekhattar R . (2003). A candidate X-linked mental retardation gene is a component of a new family of histone deacetylase-containing complexes. J Biol Chem 278: 7234–7239.

    Article  CAS  Google Scholar 

  • Hamamoto R, Furukawa Y, Morita M, Iimura Y, Silva FP, Li M et al. (2004). SMYD3 encodes a histone methyltransferase involved in the proliferation of cancer cells. Nat Cell Biol 6: 731–740.

    Article  CAS  Google Scholar 

  • Huang G, Krig S, Kowbel D, Xu H, Hyun B, Volik S et al. (2005). ZNF217 suppresses cell death associated with chemotherapy and telomere dysfunction. Hum Mol Genet 14: 3219–3225.

    Article  CAS  Google Scholar 

  • Iuchi S . (2001). Three classes of C2H2 zinc finger proteins. Cell Mol Life Sci 58: 625–635.

    Article  CAS  Google Scholar 

  • Lee MG, Wynder C, Cooch N, Shiekhattar R . (2005). An essential role for CoREST in nucleosomal histone 3 lysine 4 demethylation. Nature 437: 432–435.

    Article  CAS  Google Scholar 

  • Metzger D, Wisseann M, Yin N, Muller JM, Schneider R, Peters AH et al. (2005). LSD1 demethylates repressive histone marks to promote androgen-receptor-dependent transcription. Nature 437: 436–439.

    Article  CAS  Google Scholar 

  • Nishioka K, Chuikov S, Sarma K, Erdjument-Bromage H, Allis CD, Tempst P et al. (2002). Set9, a novel histone H3 methyltransferase that facilitates transcription by precluding histone tail modifications required for heterochromatin formation. Genes Dev 16: 479–489.

    Article  CAS  Google Scholar 

  • Perez-Moreno MA, Locascio A, Rodrigo I, Dhondt G, Portillo F, Nieto MA et al. (2001). A new role for E12/E47 in the repression of E-cadherin expression and epithelial–mesenchymal transitions. J Biol Chem 276: 27424–27431.

    Article  CAS  Google Scholar 

  • Quinlan KG, Nardini M, Verger A, Francescato P, Yaswen P, Corda D et al. (2006). Specific recognition of ZNF217 and other zinc-finger proteins at a surface groove of CtBPs. Mol Cell Biol 26: 8159–8172.

    Article  CAS  Google Scholar 

  • Shi Y, Sawada J, Sui G, Affar el B, Whetstine JR, Lan F et al. (2003). Coordinated histone modifications mediated by a CtBP co-repressor complex. Nature 422: 735–738.

    Article  CAS  Google Scholar 

  • Shi Y, Lan F, Matson C, Mulligan P, Whetstine JR, Cole PA et al. (2004). Histone demethylation mediated by the nuclear amine oxidase homolog LSD1. Cell 119: 941–953.

    Article  CAS  Google Scholar 

  • Takeichi M . (1993). Cadherins in cancer: implications for invasion and metastasis. Curr Opin Cell Biol 5: 806–811.

    Article  CAS  Google Scholar 

  • Tanner MM, Tirkkonen M, Kallioniemi A, Collins C, Stokke T, Karhu R et al. (1994). Increased copy number at 20q13 in breast cancer: defining the critical region and exclusion of candidate genes. Cancer Res 54: 4257–4260.

    CAS  PubMed  Google Scholar 

  • Tanner MM, Tirkkonen M, Kallioniemi A, Holli K, Collins C, Kowbel D et al. (1995). Amplification of chromosomal region 20q13 in invasive breast cancer: prognostic implications. Clin Cancer Res 1: 1455–1461.

    CAS  PubMed  Google Scholar 

  • Thiery JP . (2002). Epithelial–mesenchymal transitions in tumour progression. Nat Rev Cancer 2: 442–454.

    Article  CAS  Google Scholar 

  • Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C et al. (2004). Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117: 927–939.

    Article  CAS  Google Scholar 

  • You A, Tong JK, Grozinger CM, Schreiber SL . (2001). CoREST is an integral component of the CoREST–human histone deacetylase complex. Proc Natl Acad Sci USA 98: 1454–1458.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Gail Mandel for providing the HIS-tagged CoREST cDNA and Paul Yaswen for providing the HA-tagged ZNF217. We acknowledge the technical assistance of Melissa Coubrough and Thu Tran. This research was supported by grants from the Cancer Research Society of Canada and the Lawson Health Research Foundation. JJM Cowger is a recipient of a predoctoral fellowship from The Natural Science and Engineering Research Council of Canada (NSERC).

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Authors and Affiliations

  1. Department of Oncology, London Regional Cancer Program, The University of Western Ontario, London, Ontario, Canada

    J J M Cowger, Q Zhao, M Isovic & J Torchia

  2. Department of Physiology and Pharmacology, The University of Western Ontario, London, Ontario, Canada

    J J M Cowger & J Torchia

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  1. J J M Cowger
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  2. Q Zhao
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  3. M Isovic
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  4. J Torchia
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Correspondence to J Torchia.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Cowger, J., Zhao, Q., Isovic, M. et al. Biochemical characterization of the zinc-finger protein 217 transcriptional repressor complex: identification of a ZNF217 consensus recognition sequence. Oncogene 26, 3378–3386 (2007). https://doi.org/10.1038/sj.onc.1210126

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