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ETV6/RUNX1 abrogates mitotic checkpoint function and targets its key player MAD2L1

Oncogene volume 29, pages 3307–3312 (2010)Cite this article

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Abstract

Approximately 25% of childhood B-cell precursor acute lymphoblastic leukemia have an ETV6/RUNX1 (E/R) gene fusion that results from a t(12;21). This genetic subgroup of leukemia is associated with near-triploidy, near-tetraploidy, and trisomy 21 as rather specific types of secondary changes. Here, we show that, unlike various controls, E/R-expressing Ba/F3 clones acquire a tetraploid karyotype on prolonged culture, corroborating the assumption that E/R may attenuate the mitotic checkpoint (MC). Consistent with this notion, E/R-expressing diploid murine and human cell lines have decreased proportions of cells with 4N DNA content and a lower mitotic index when treated with spindle toxins. Moreover, both RUNX1 and E/R regulate mitotic arrest-deficient 2 L1 (MAD2L1), an essential MC component, by binding to promoter-inherent RUNX1 sites, which results in down-regulation of MAD2L1 mRNA and protein in E/R-expressing cells. Forced expression of E/R also abolishes RUNX1-induced reporter activation, whereas E/R with a mutant DNA-binding site leads to only minor effects. Our data link for the first time E/R, MC, and MAD2L1 and provide new insights into the function of the E/R fusion gene product. Although tetraploidy is an almost exclusive feature of E/R-positive leukemias, its rarity within this particular subgroup implies that further yet unknown factors are required for its manifestation.

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References

  • Attarbaschi A, Mann G, Konig M, Dworzak MN, Trebo MM, Muhlegger N et al. (2004). Incidence and relevance of secondary chromosome abnormalities in childhood TEL/AML1+ acute lymphoblastic leukemia: an interphase FISH analysis. Leukemia 18: 1611–1616.

    Article  CAS  Google Scholar 

  • Attarbaschi A, Mann G, Konig M, Steiner M, Dworzak MN, Gadner H et al. (2006). Near-tetraploidy in childhood B-cell precursor acute lymphoblastic leukemia is a highly specific feature of ETV6/RUNX1-positive leukemic cases. Genes Chromosomes Cancer 45: 608–611.

    Article  CAS  Google Scholar 

  • Boyapati A, Yan M, Peterson LF, Biggs JR, Le Beau MM, Zhang DE . (2007). A leukemia fusion protein attenuates the spindle checkpoint and promotes aneuploidy. Blood 109: 3963–3971.

    Article  CAS  Google Scholar 

  • Diakos C, Krapf G, Gerner C, Inthal A, Lemberger C, Ban J et al. (2007). RNAi-mediated silencing of TEL/AML1 reveals a heat-shock protein- and survivin-dependent mechanism for survival. Blood 109: 2607–2610.

    Article  CAS  Google Scholar 

  • Fang Y, Liu T, Wang X, Yang YM, Deng H, Kunicki J et al. (2006). BubR1 is involved in regulation of DNA damage responses. Oncogene 25: 3598–3605.

    Article  CAS  Google Scholar 

  • Ford AM, Palmi C, Bueno C, Hong D, Cardus P, Knight D et al. (2009). The TEL-AML1 leukemia fusion gene dysregulates the TGF-beta pathway in early B lineage progenitor cells. J Clin Invest 119: 826–836.

    CAS  Google Scholar 

  • Friedman AD . (1999). Leukemogenesis by CBF oncoproteins. Leukemia 13: 1932–1942.

    Article  CAS  Google Scholar 

  • Guardavaccaro D, Frescas D, Dorrello NV, Peschiaroli A, Multani AS, Cardozo T et al. (2008). Control of chromosome stability by the beta-TrCP-REST-Mad2 axis. Nature 452: 365–369.

    Article  CAS  Google Scholar 

  • Ha GH, Baek KH, Kim HS, Jeong SJ, Kim CM, McKeon F et al. (2007). p53 activation in response to mitotic spindle damage requires signaling via BubR1-mediated phosphorylation. Cancer Res 67: 7155–7164.

    Article  CAS  Google Scholar 

  • Hiebert SW, Sun W, Davis JN, Golub T, Shurtleff S, Buijs A et al. (1996). The t(12;21) translocation converts AML-1B from an activator to a repressor of transcription. Mol Cell Biol 16: 1349–1355.

    Article  CAS  Google Scholar 

  • Hong D, Gupta R, Ancliff P, Atzberger A, Brown J, Soneji S et al. (2008). Initiating and cancer-propagating cells in TEL-AML1-associated childhood leukemia. Science 319: 336–339.

    Article  CAS  Google Scholar 

  • Kops GJ, Weaver BA, Cleveland DW . (2005). On the road to cancer: aneuploidy and the mitotic checkpoint. Nat Rev Cancer 5: 773–785.

    Article  CAS  Google Scholar 

  • Li Z, Yan J, Matheny CJ, Corpora T, Bravo J, Warren AJ et al. (2003). Energetic contribution of residues in the Runx1 Runt domain to DNA binding. J Biol Chem 278: 33088–33096.

    Article  CAS  Google Scholar 

  • Lou J, Cao W, Bernardin F, Ayyanathan K, Rauscher IF, Friedman AD . (2000). Exogenous cdk4 overcomes reduced cdk4 RNA and inhibition of G1 progression in hematopoietic cells expressing a dominant-negative CBF—a model for overcoming inhibition of proliferation by CBF oncoproteins. Oncogene 19: 2695–2703.

    Article  CAS  Google Scholar 

  • Michel L, Benezra R, Diaz-Rodriguez E . (2004). MAD2 dependent mitotic checkpoint defects in tumorigenesis and tumor cell death: a double edged sword. Cell Cycle 3: 990–992.

    Article  CAS  Google Scholar 

  • Morrow M, Samanta A, Kioussis D, Brady HJ, Williams O . (2007). TEL-AML1 preleukemic activity requires the DNA binding domain of AML1 and the dimerization and corepressor binding domains of TEL. Oncogene 26: 4404–4414.

    Article  CAS  Google Scholar 

  • Perez de Castro I, de Carcer G, Malumbres M . (2007). A census of mitotic cancer genes: new insights into tumor cell biology and cancer therapy. Carcinogenesis 28: 899–912.

    Article  CAS  Google Scholar 

  • Peters JM . (2007). Cell biology: the checkpoint brake relieved. Nature 446: 868–869.

    Article  CAS  Google Scholar 

  • Pines J . (2006). Mitosis: a matter of getting rid of the right protein at the right time. Trends Cell Biol 16: 55–63.

    Article  CAS  Google Scholar 

  • Pui CH, Relling MV, Downing JR . (2004). Acute lymphoblastic leukemia. N Engl J Med 350: 1535–1548.

    Article  CAS  Google Scholar 

  • Raimondi SC, Zhou Y, Shurtleff SA, Rubnitz JE, Pui CH, Behm FG . (2006). Near-triploidy and near-tetraploidy in childhood acute lymphoblastic leukemia: association with B-lineage blast cells carrying the ETV6-RUNX1 fusion, T-lineage immunophenotype, and favorable outcome. Cancer Genet Cytogenet 169: 50–57.

    Article  CAS  Google Scholar 

  • Ross ME, Zhou X, Song G, Shurtleff SA, Girtman K, Williams WK et al. (2003). Classification of pediatric acute lymphoblastic leukemia by gene expression profiling. Blood 102: 2951–2959.

    Article  CAS  Google Scholar 

  • Roudaia L, Cheney MD, Manuylova E, Chen W, Morrow M, Park S et al. (2009). CBFbeta is critical for AML1-ETO and TEL-AML1 activity. Blood 113: 3070–3079.

    Article  CAS  Google Scholar 

  • Sasaki K, Nakamura Y, Maki K, Waga K, Nakamura F, Arai H et al. (2004). Functional analysis of a dominant-negative DeltaETS TEL/ETV6 isoform. Biochem Biophys Res Commun 317: 1128–1137.

    Article  CAS  Google Scholar 

  • Song WJ, Sullivan MG, Legare RD, Hutchings S, Tan X, Kufrin D et al. (1999). Haploinsufficiency of CBFA2 causes familial thrombocytopenia with propensity to develop acute myelogenous leukaemia. Nat Genet 23: 166–175.

    Article  CAS  Google Scholar 

  • Sotillo R, Hernando E, Diaz-Rodriguez E, Teruya-Feldstein J, Cordon-Cardo C, Lowe SW et al. (2007). Mad2 overexpression promotes aneuploidy and tumorigenesis in mice. Cancer Cell 11: 9–23.

    Article  CAS  Google Scholar 

  • Speck NA, Gilliland DG . (2002). Core-binding factors in haematopoiesis and leukaemia. Nat Rev Cancer 2: 502–513.

    Article  CAS  Google Scholar 

  • Strom DK, Nip J, Westendorf JJ, Linggi B, Lutterbach B, Downing JR et al. (2000). Expression of the AML-1 oncogene shortens the G(1) phase of the cell cycle. J Biol Chem 275: 3438–3445.

    Article  CAS  Google Scholar 

  • Weaver BA, Cleveland DW . (2005). Decoding the links between mitosis, cancer, and chemotherapy: the mitotic checkpoint, adaptation, and cell death. Cancer Cell 8: 7–12.

    Article  CAS  Google Scholar 

  • Wolyniec K, Wotton S, Kilbey A, Jenkins A, Terry A, Peters G et al. (2009). RUNX1 and its fusion oncoprotein derivative, RUNX1-ETO, induce senescence-like growth arrest independently of replicative stress. Oncogene 28: 2502–2512.

    Article  CAS  Google Scholar 

  • Wotton S, Terry A, Kilbey A, Jenkins A, Herzyk P, Cameron E et al. (2008). Gene array analysis reveals a common Runx transcriptional programme controlling cell adhesion and survival. Oncogene 27: 5856–5866.

    Article  CAS  Google Scholar 

  • Zelent A, Greaves M, Enver T . (2004). Role of the TEL-AML1 fusion gene in the molecular pathogenesis of childhood acute lymphoblastic leukaemia. Oncogene 23: 4275–4283.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank O Williams for expression vectors containing E/R or the mutant E/R, D-E Zhang for the RUNX1-expression vector, A Friedman for CBFβ-expression vector, D Guardavaccaro for the luciferase reporter plasmid for the MAD2L1 promoter in pGL3, JD Rowley for AT-2 cell line, L Orel for the Myc-ETV6/RUNX1 vector, Idriss M Bennani-Baiti for stimulating discussions, and Marion Zavadil for proofreading the paper. This study was supported in part by a grant from the FWF P17551-B14 and the Austrian Ministry for Education, Science and Culture (GENAU-Ch.I.L.D) and the St Anna Kinderkrebsforschung to ER P-G. A Kilbey is funded by the Leukaemia Research Fund. GK participated in the design of the study, performed experiments, interpreted data, and wrote the paper. UK, GF, AI, RJ performed research and interpreted data. AK performed and interpreted ChIP experiments. JCN participated in study design and interpretation of ChIP data. GM contributed patient samples. OAH participated in the interpretation of data and writing of the paper. ER P-G designed and supervised research and wrote the paper.

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Authors and Affiliations

  1. Children's Cancer Research Institute, Vienna, Austria

    G Krapf, U Kaindl, G Fuka, A Inthal, R Joas & E R Panzer-Grümayer

  2. Molecular Oncology Laboratory, Institute of Comparative Medicine, Faculty of Veterinary Medicine, Glasgow, UK

    A Kilbey & J C Neil

  3. St Anna Kinderspital, Vienna, Austria

    G Mann, O A Haas & E R Panzer-Grümayer

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  1. G Krapf
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  2. U Kaindl
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  3. A Kilbey
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  4. G Fuka
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Correspondence to E R Panzer-Grümayer.

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Krapf, G., Kaindl, U., Kilbey, A. et al. ETV6/RUNX1 abrogates mitotic checkpoint function and targets its key player MAD2L1. Oncogene 29, 3307–3312 (2010). https://doi.org/10.1038/onc.2010.53

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