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J. Biol. Chem., Vol. 275, Issue 33, 25292-25298, August 18, 2000

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Ectopic Expression of Transcription Factor NF-E2 Alters the Phenotype of Erythroid and Monoblastoid Cells^*

Melissa S. Sayer^§, Peta A. Tilbrook^§, Angelo Spadaccini^§, Evan Ingley^§, Mohinder K. Sarna^§, James H. Williams^§, Nancy C. Andrews^¶, and S. Peter Klinken^§

From the  Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital, Perth WA 6000, Australia, the ^§ Department of Biochemistry, University of Western Australia, Nedlands 6907, Australia, and the ^¶ Howard Hughes Medical Institute, Children's Hospital, and Harvard Medical School, Boston, Massachusetts 02115

In this study, regulation of transcription factor NF-E2 was examined in differentiating erythroid and myeloid cells, and the impact of raising NF-E2 concentrations within these cell types was assessed. NF-E2 was expressed in the J2E erythroid cell line, but the levels increased only marginally during erythropoietin-induced differentiation. In contrast, rare myeloid variants of J2E cells did not express NF-E2. Although NF-E2 was present in M1 monoblastoid cells, it was undetectable as these cells matured into macrophages. Compared with erythroid cells, transcription of the NF-E2 gene was reduced, and the half-life of the mRNA was significantly shorter in monocytoid cells. Ectopic expression of NF-E2 had a profound impact upon the J2E cells; morphologically mature erythroid cells spontaneously emerged in culture, but the cells failed to synthesize hemoglobin, even in the presence of erythropoietin. Although proliferation and viability increased in the NF-E2-transfected J2E cells, their responsiveness to erythropoietin was severely diminished. Strikingly, increasing the expression of NF-E2 in M1 cells produced sublines that contained erythroid or immature megakaryocytic cells. Finally, overexpression of NF-E2 in primary hemopoietic progenitors from fetal liver increased erythroid colony formation in the absence of erythropoietin. These data demonstrate that elevated NF-E2 (i) had a dominant effect on the phenotype and maturation of J2E erythroid cells, (ii) was able to reprogram the M1 monocytoid line, and (iii) promoted the development of erythroid colonies by normal progenitors.

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