Comparative Proteomic Analysis of Human Leukemic Cells with and without Inducible Expression of Leukemogenic AML1-ETO Protein

Lei Zhang；Li-Shun Wang；Ying Xu；Li Xia；Wen-Li Chen；Ying Zheng；Guo-Qiang Chen

leukemia ； AML1-ETO ； proteomics ； MORG1 ； MAT1

The Chinese Journal of Physiology

49卷4期（2006 / 08 / 31）

182 - 191

英文

AML1-ETO is a leukemogenic fusion protein generated by chromosomal translocation t(8; 21) (q22; q22), one of the most frequent chromosomal abnormalities in acute myeloid leukemia. The fusion protein has been shown to present dichotomous functions on leukemic cells: growth arrest versus differentiation block. However, their precise molecular mechanisms are not completely clear. In this work, we try to explore potential AML1-ETO-targeted proteins through comparing two-dimensional electrophoresis (2DE)-based global protein expression profiles of leukemic U937 cells with and without inducible expression of AML1-ETO. As a result, we identified 14 unique proteins deregulated in AML1-ETO-carrying leukemic cells, including 3 up-regulated such as hairy and enhancer of split 5 (HES5) and 11 down-regulated such as MAT1 (menage a trois-1) and mitogen-activated protein kinase organizer 1 (MORG1). These proteins were widely involved in stem cell maintenance, cell cycle, signal transduction and transcription. The further investigation on their roles in leukemic cells will uncover new clues to understanding leukemogenic effects of AML1-ETO fusion protein.

1. Ahn, M.Y.,Huang, G.,Bae, S.C.,Wee, H.J.,Kim, W.Y.,Ito, Y.(1998).Negative regulation of granulocytic differentiation in the myeloid precursor cell line 32Dcl3 by ear-2, a mammalian homolog of Drosophila seven-up, and a chimeric leukemogenic gene, AML1/ ETO.Proc. Natl. Acad. Sci. USA,95,1812-1817.
2. Alcalay, M.,Meani, N.,Gelmetti, V.,Fantozzi, A.,Fagioli, M.,Orleth, A.,Riganelli, D.,Sebastiani, C.,Cappelli, E.,Casciari, C.,Sciurpi, M.T.,Mariano, A.R.,Minardi, S.P.,Luzi, L.,Muller, H.,Di Fiore, P.P.,Frosina, G.,Pelicci, P.G.(2003).Acute myeloid leukemia fusion proteins deregulate genes involved in stem cell maintenance and DNA repair.J. Clin. Invest,112,1751-1761.
3. Amann, J.M.,Nip, J.,Strom, D.K.,Lutterbach, B.,Harada, H.,Lenny, N.,Downing, J.R.,Meyers, S.,Hiebert, S.W.(2001).ETO, a target of t(8;21) in acute leukemia, makes distinct contacts with multiple histone deacetylases and binds mSin3A through its oligomerization domain.Mol. Cell. Biol,21,6470-6483.
4. Behre, G.,Zhang, P.,Zhang, D.E.,Tenen, D.G.(1999).Analysis of the modulation of transcriptional activity in myelopoiesis and leukemogenesis.Methods,17,231-237.
5. Biggs, J.R.,Zhang, Y.,Peterson, L.F.,Garcia, M.,Zhang, D.E.,Kraft, A.S.(2005).Phosphorylation of AML1/RUNX1 regulates its degradation and nuclear matrix association.Mol. Cancer Res,3,391-401.
6. Burel, S.A.,Harakawa, N.,Zhou, L.,Pabst, T.,Tenen, D.G.,Zhang, D.E.(2001).Dichotomy of AML1-ETO functions: growth arrest versus block of differentiation.Mol. Cell. Biol,21,5577-5590.
7. Chiaramonte, R.,Basile, A.,Tassi, E.,Calzavara, E.,Cecchinato, V.,Rossi, V.,Biondi, A.,Comi, P.(2005).A wide role for NOTCH1 signaling in acute leukemia.Cancer Lett,219,113-120.
8. Fliegauf, M.,Stock, M.,Berg, T.,Lubbert, M.(2004).Williams-Beuren syndrome critical region-5/non-T-cell activation linker: a novel target gene of AML1/ETO.Oncogene,23,9070-9081.
9. Giglia-Mari, G.,Coin, F.,Ranish, J.A.,Hoogstraten, D.,Theil, A.,Wijgers, N.,Jaspers, N.G.,Raams, A.,Argentini, M.,van der Spek, P.J.,Botta, E.,Stefanini, M.,Egly, J.M.,Aebersold, R.,Hoeijmakers, J.H.,Vermeulen, W.(2004).A new, tenth subunit of TFIIH is responsible for the DNA repair syndrome trichothiodystrophy group A.Nat. Genet,36,714-719.
10. Heidenreich, O.,Krauter, J.,Riehle, H.,Hadwiger, P.,John, M.,Heil, G.,Vornlocher, H.P.,Nordheim, A.(2003).AML1/MTG8 oncogene suppression by small interfering RNAs supports myeloid differentiation of t(8;21)-positive leukemic cells.Blood,101,3157-3163.
11. Hiebert, S.W.,Lutterbach, B.,Amann, J.(2001).Role of co-repressors in transcriptional repression mediated by the t(8;21), t(16;21), t(12; 21), and inv(16) fusion proteins.Curr. Opin. Hematol,8,197-200.
12. Jiang, Y.,Xue, Z.H.,Shen, W.Z.,Du, K.M.,Yan, H.,Yu, Y.,Peng, Z.G.,Song, M.G.,Tong, J.H.,Chen, Z.,Huang, Y.,Lubbert, M.,Chen, G.Q.(2005).Desferrioxamine induces leukemic cell differentiation potentially by hypoxia-inducible factor-1 alpha that augments transcriptional activity of CCAAT/enhancer-binding protein-alpha.Leukemia,19,1239-1247.
13. Kurokawa, M.,Hirai, H.(2003).Role of AML1/Runx1 in the pathogenesis of hematological malignancies.Cancer Sci,94,841-846.
14. Kurokawa, M.,Tanaka, T.,Tanaka, K.,Ogawa, S.,Mitani, K.,Yazaki, Y.,Hirai, H.(1996).Overexpression of the AML1 protooncoprotein in NIH3T3 cells leads to neoplastic transformation depending on the DNA-binding and transactivational potencies.Oncogene,12,883-892.
15. Look, A.T.(1997).Oncogenic transcription factors in the human acute leukemias.Science,278,1059-1064.
16. Lutterbach, B.,Westendorf, J.J.,Linggi, B.,Patten, A.,Moniwa, M.,Davie, J.R.,Huynh, K.D.,Bardwell, V.J.,Lavinsky, R.M.,Rosenfeld, M.G.,Glass, C.,Seto, E.,Hiebert, S.W.(1998).ETO, a target of t(8;21) in acute leukemia, interacts with the N-CoR and mSin3 corepressors.Mol. Cell. Biol,18,7176-7184.
17. Mao, S.,Frank, R.C.,Zhang, J.,Miyazaki, Y.,Nimer, S.D.(1999).Functional and physical interactions between AML1 proteins and an ETS protein, MEF: implications for the pathogenesis of t(8;21)- positive leukemias.Mol. Cell. Biol,19,3635-3644.
18. Mori, H.,Colman, S.M.,Xiao, Z.,Ford, A.M.,Healy, L.E.,Donaldson, C.,Hows, J.M.,Navarrete, C.,Greaves, M.(2002).Chromosome translocations and covert leukemic clones are generated during normal fetal development.Proc. Natl. Acad. Sci. USA,99,8242-8247.
19. Okuda, T.,van Deursen, J.,Hiebert, S.W.,Grosveld, G.,Downing, J.R.(1996).AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis.Cell,84,321-330.
20. Peterson, L.F.,Zhang, D.E.(2004).The 8;21 translocation in leukemogenesis.Oncogene,23,4255-4262.
21. Puccetti, E.,Obradovic, D.,Beissert, T.,Bianchini, A.,Washburn, B.,Chiaradonna, F.,Boehrer, S.,Hoelzer, D.,Ottmann, O.G.,Pelicci, P.G.,Nervi, C.,Ruthardt, M.(2002).AML-associated translocation products block vitamin D(3)-induced differentiation by sequestering the vitamin D(3) receptor.Cancer Res,62,7050-7058.
22. Rhoades, K.L.,Hetherington, C.J.,Rowley, J.D.,Hiebert, S.W.,Nucifora, G.,Tenen, D.G.,Zhang, D.E.(1996).Synergistic up-regulation of the myeloid-specific promoter for the macrophage colonystimulating factor receptor by AML1 and the t(8;21) fusion protein may contribute to leukemogenesis.Proc. Natl. Acad. Sci. USA,93,11895-11900.
23. Schwieger, M.,Lohler, J.,Friel, J.,Scheller, M.,Horak, I.,Stocking, C.(2002).AML1-ETO inhibits maturation of multiple lymphohematopoietic lineages and induces myeloblast transformation in synergy with ICSBP deficiency.J. Exp. Med,196,1227-1240.
24. Shimada, H.,Ichikawa, H.,Ohki, M.(2002).Potential involvement of the AML1-MTG8 fusion protein in the granulocytic maturation characteristic of the t(8;21) acute myelogenous leukemia revealed by microarray analysis.Leukemia,16,874-885.
25. Steffen, B.,Serve, H.,Berdel, W.E.,Agrawal, S.,Linggi, B.,Buchner, T.,Hiebert, S.W.,Muller-Tidow, C.(2003).Specific protein redirection as a transcriptional therapy approach for t(8;21) leukemia.Proc. Natl. Acad. Sci. USA,100,8448-8453.
26. Tanaka, K.,Tanaka, T.,Kurokawa, M.,Imai, Y.,Ogawa, S.,Mitani, K.,Yazaki, Y.,Hirai, H.(1998).The AML1/ETO(MTG8) and AML1/Evi-1 leukemia-associated chimeric oncoproteins accumulate PEBP2beta(CBFbeta) in the nucleus more efficiently than wildtype AML1.Blood,91,1688-1699.
27. Tanaka, T.,Kurokawa, M.,Ueki, K.,Tanaka, K.,Imai, Y.,Mitani, K.,Okazaki, K.,Sagata, N.,Yazaki, Y.,Shibata, Y.,Kadowaki, T.,Hirai, H.(1996).The extracellular signal-regulated kinase pathway phosphorylates AML1, an acute myeloid leukemia gene product, and potentially regulates its transactivation ability.Mol. Cell. Biol,16,3967-3979.
28. Tassan, J.P.,Jaquenoud, M.,Fry, A.M.,Frutiger, S.,Hughes, G.J.,Nigg, E.A.(1995).In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein.EMBO. J.,14,5608-5617.
29. Tenen, D.G.(2003).Disruption of differentiation in human cancer: AML shows the way.Nat. Rev. Cancer,3,89-101.
30. Tenen, D.G.,Hromas, R.,Licht, J.D.,Zhang, D.E.(1997).Transcription factors, normal myeloid development, and leukemia.Blood,90,489-519.
31. Vangala, R.K.,Heiss-Neumann, M.S.,Rangatia, J.S.,Singh, S.M.,Schoch, C.,Tenen, D.G.,Hiddemann, W.,Behre, G.(2003).The myeloid master regulator transcription factor PU.1 is inactivated by AML1-ETO in t(8;21) myeloid leukemia.Blood,101,270-277.
32. Vomastek, T.,Schaeffer, H.J.,Tarcsafalvi, A.,Smolkin, M.E.,Bissonette, E.A.,Weber, M.J.(2004).Modular construction of a signaling scaffold: MORG1 interacts with components of the ERK cascade and links ERK signaling to specific agonists.Proc. Natl. Acad. Sci. USA,101,6981-6986.
33. Wiemels, J.L.,Xiao, Z.,Buffler, P.A.,Maia, A.T.,Ma, X.,Dicks, B.M.,Smith, M.T.,Zhang, L.,Feusner, J.,Wiencke, J.,Pritchard- Jones, K.,Kempski, H.,Greaves, M.(2002).In utero origin of t(8;21) AML1-ETO translocations in childhood acute myeloid leukemia.Blood,99,3801-3805.