Abstract
In this study, we analyze the binding of nuclear proteins isolated from the hydroxyurea (Hu)-induced and uninduced HEL cells to the DNasel hypersensitive sites III (HS3-14991-14716 bp) and IV(HS4 -18586-18306 bp) in the human β-globin gene locus control region (LCR). Using Western blot assay, we demonstrate that GATA-1 transcription factor in HEL cells is increased following the induction of Hu, while GATA-2 transcription factor is decreased. Based on both the competition EMSA and the Western blot assay, our data reveal that the nuclear protein isolated from the uninduced HEL cells, which can bind to the HS3 and HS4 core DNA sequences, is mainly GATA-2; however, the nuclear proteins isolated from the Hu-induced HEL cells, which can bind to the HS3 and HS4 core DNA sequences, are mainly GATA-1 and GATA-X (a kind of unknown GATA factor). These results suggest that the erythroid specific transcription factors (GATA-1 and GATA-2) in the Hu-induced and uninduced HEL cells can selectively bind to the HS3 and HS4 core DNA sequences in the human β-globin gene LCR, indicating that GATA factors (GATA-1 and GATA-2) may play very important role in the regulation of the β-globin gene expression.
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References
Marin, P., Papayannopoulou, T., HEL cells: a new erythroleukemia cell line with spontaneous and induced globin expression, Science. 1982, 216(4551): 1233.
Rodgere, G. P., Dover, G. J., Noguchi, C. T. et al., Hematologic responses of patients with sickle cell disease to treatment with hydroxyurea, N. Engl. J. Med., 1990, 322(15): 1037.
Huang, S. Z., Ren, B. R., Chen, M. Y. et al., Treatment of β-thalassemia with hydroxyurea (Hu), Science in China, Ser, B, 1994, 37(11): 1350.
Gui, C. Y., Jiang, C., Qian, R. L., The role of hydroxyurea in globin gene expression and cell differentiation of human erythroleukemia cell line, Acta Biologiac Experimentalis Sinica, 1997, 30(4): 375.
Felsonfeld, G., Chromatin as an essential part of transcriptional mechanism, Nature, 1992, 355(6357): 219.
Orkin, S. H., Regulation of globin gene expression in erythroid cells, Eur. J. Biochem., 1995, 231(2): 271.
Fraser, P., Pruzina, S., Antoniou, M. et al., Each hypersensitive site of the human β-globin locus control region confers a different developmental pattern of expression of the globin genes, Gene & Dev., 1993, 7(1): 106.
Dignam, J. D., Lebovitz, R. M., Roeder, R. G., Accurate transcription initiation by RNA polymerasell in a soluble extract from isolated mammalian nuclei, Nucleic Acids Res., 1983, 11(5): 1475.
Ausubel, F. M., Brent, R., Kingston, R. E., et al., Preparation of nuclear and cytoplasmic extracts from mammalian cells. Short Protocols in Molecular Biology, 2nd ed., New York: John Wiley & Sons Press, 1992.
Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 1976, 72(1): 248.
Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.
Yan, Z. J., Chen, Y. D., Qian, R. L., Developmental stage-specific factors in the mouse hematopoietic tissues binding to the 5′-flanking cis-acting elements of human β-globin gene, Chinese Science Bulletin, 1995, 40(9): 778.
Orkin, S. H., GATA-binding transcription factors in hematopoietic cells, Blood, 1992, 80(2): 575.
Philipsen, S., Titlbot, D., Flaser, P. et al., The β-globin dominant control region: hypersensitive site 2, The EMBO Journal, 1990. 9(7): 2159.
Pruzina, S., Olivia, H., Grosveld, F. et al., Hypersensitive site 4 of the human β-globin locus control region, Nucleic Acids Res., 1991, 19(7): 1413.
Wijgerde, M., Grosveld, F., Fraser, P., Transcription complex stability and chromatin dynamicsin vivo, Nature, 1995, 377(6546): 209.
Stamatoyannopoulos, J. A., Goodwin, A., Joyce, T. et al., NF-E2 and GATA binding motif are required for the formation of DNaseI hypersensitive site 4 of the human β-globin locus control region, The EMBO Journal, 1995, 14(1): 106.
Boulitas, T., Homeodomain protein binding sites, inverted repeats, and nuclear matrix attachment regions along the human β-globin gene complex. Journal of Cellular Biochemistry, 1993, 52(1): 23.
Persons, D. A., Allay, J. A., Nienhuis, A. W., Enforced expression of GATA-2 transcription factor blocks normal hematopoiesis, Blood, 1999, 93(2): 488.
Fujiwara, Y., Browne, C. P., Orkin, S. H. et al., Arrested development of embryonic red cell precursors in mouse embryos lacking transcription factor GATA-1, Proc. Nat. Acad. Sci. USA, 1996, 93(22): 12355.
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Zhang, S., Zhao, H., Jiang, S. et al. Interactions between GATA transcription factors and the DNaseI hypersensitive sites in the human β-globin gene LCR. Chin.Sci.Bull. 45, 1969–1973 (2000). https://doi.org/10.1007/BF02909689
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DOI: https://doi.org/10.1007/BF02909689