Abstract
The induction of differentiation in mouse erythroleukemia (MEL) cells by dimethylosulfoxide (DMSO) is characterized by increased transcription of globin genes. We have determined that DMSO treated cells increase the levels of nuclear factors capable of overall interactions with the βmaj globin promoter during the initial 24 h post induction, as measured by gel mobility analysis. Two unprocessed βmaj globin mRNA precursors, which are present in MEL cell nuclei early in differentiation, were previously shown to contain the 5′ promoter flanking region, and thereby provided the nucleus with a pool of regulatory sequences in multiple RNA copies. We have studied the effect of RNA copies of the promoter region on binding interactions between DNA sequences of the βmaj globin promoter and nuclear factors that interact with these sequences. The promoter region RNA transcripts competed effectively for DNA binding proteins in vitro, while the antisense RNA from the same region did not. The most pronounced competition was observed with proteins from 12 h after DMSO induction, when the concentration of the DNA binding proteins was still increasing. Since the ‘upstream’ transcripts predominate at 12 h after DMSO induction, these results indicate that the promoter region transcripts may influence the equilibrium of binding between the βmaj globin promoter an the nuclear factors that bind to this region during DMSO induction.
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References
Evans T, Felsenfeld G, Reitman M: Control of globin gene transcription. Annu Rev Cell Biol 6:95–124, 1990
Orkin S: Globin gene regulation and switching: circa 1990. Cel 63: 665–672, 1990
Benezra R, Cantor RC, Axel R: Nucleosomes are phased along the mouse β-major globin gene in erythroid and nonerythroid cells. Cell 44:697–704, 1986
Charnay P, Mellon P, Maniatis T: Linker scanning mutagenesis of the 5′-flanking region of mouse β-major globin gene: sequence requirements for transcription in erythroid and nonerythroid cells. Mol Cell Biol 5:1498–1511, 1985
Cowie A, Myers RM: DNA sequences involved in transcriptional regulation of the mouse β-globin promoter in murine erythroleukemia cells. Mol Cell Biol 8:3122–3128, 1988
Myers RM, Tilly K, Maniatis T: Fine structure genetic analysis of β-globin promoter. Science 232:613–618, 1986
Myers RM, Cowie A, Stuve L, Hartzog G, Gaensler K: Genetic and biochemical analysis of the mouse β-major globin promoter. In: G. Stamatoyannopoulos and A.W. Nienhuis (eds). Hemoglobin Switching Part A: Transcriptional Regulation. A.R. Liss Inc, New York, 1989, pp 117–127
Stuve LL, Myers RM: A directly repeated sequence in the β-globin promoter regulates transcription in murine erythroleukemia cells. Mol Cell Biol 10:972–981, 1990
Hartzog GA, Myers RM: Discrimination among potential activators of the β-globin CACCC element by correlation of binding and transcriptional properties. Mol Cell Biol 13:44–56, 1993
Stuve LL, Myers RM: Identification and characterization of a β-globin promoter-binding factor from murine erythroleukemia cells. Mol Cell Biol 13:4311–4322, 1993
deBoer E, Antoniou M, Mignotte V, Wall L, Grosveld F: The human β-globin promoter; nuclear protein factors and erythroid specific induction transcription. EMBO J 7:4203–4212, 1988
Evans T, Reitman M, Felsenfeld G: An erythrocyte-specific DNA-binding factor recognizes a regulatory sequence common to all chicken globin genes. Proc Natl Acad Sci USA 85:5976–5980, 1988
Barnhart KM, Kim CG, Sheffery M: Purification and characterization of an erythroid cell-specific factor that binds the murine α-and β-globin genes. Mol Cell Biol 9:2606–2614, 1989
Martin DIK, Tsai SF, Orkin SH: Increased γ-globin expression in a nondeletion HPFH mediated by an erythroid-specific DNA-binding factor. Nature (London) 338:435–438, 1989
Wall L, deBoer E, Grosveld F: The human β-globin gene 3′ enhancer contains multiple binding sites for an erythroid-specific protein. Genes Dev 2:1089–1100, 1988
Macleod K, Plumb M: Derepression of mouse β-major-globin gene transcription during erythroid differentiation. Mol Cell Biol 11: 4324–4332, 1991
Blau HM: Differentiation requires continuous active control. Annu Rev Biochem 61:1213–1230, 1992
Ley TJ, Nienhuis AW: A weak upstream promoter gives rise to long human β-globin RNA molecules. Biochem Biophys Res Commu 112: 1041–1048, 1983
Carlson DP, Ross J: Human β-globin promoter and coding sequences transcribed by RNA polymerase III. Cell 34:857–864, 1983
Allan M, Grindlay GJ, Stefani L, Paul J:Epsilon globin gene transcripts originating upstream of the mRNA capsite in K562 cells and normal human embryos. Nucleic Acids Res 10:5133–5147 1982
Allan M, Lanyon WG, Paul J: Multiple origins of transcription in the 4.5 kb upstream of the ε-globin gene. Cell 35:187–197, 1983
Carlson DP, Ross J:Alpha-amanitin-insensitive transcription of mouse βmaj-globin 5′-flanking and structural gene sequences correlates with mRNA expression. Proc Natl Acad Sci USA 81: 7782–7786, 1984
Winicov I, Weidner DA, Carlson DP, Ross J: Accuratein vitro initiation of β-globin gene transcription in induced Friend-cell nuclei. Gene 45:1–10, 1986
Weidner DA, Winicov I:Beta globin transcripts originating in the promoter region during early hexamethylene bisacetamide and dimethylsulfoxide induction of Friend erythroleukemia cells. Mol Cell Biochem 90:175–183, 1989
Frampton J, Conkie D, Chambers I, McBain W, Dexter M, Harrison P: Changes in minor transcripts from the α1 and βmaj globin and glutathione peroxidase genes during erythropoiesis. Nucleic Acids Res 15:3671–3688, 1987
Winicov I: RNA processing of β-globin transcripts containing 5′ flanking and structural gene sequences. Nucleic Acids Res 16:593–607, 1988
Carlson DP, Ross J, Wierenga PK, Nijhof W: Expression of the 5′-flanking region of the βmajor-globin gene in cultured murine erythroid precursor cells. Exp Hematol 15:902–907, 1987
Marks PA, Rifkind RA: Erythroleukemic differentiation. Ann Rev Bioch 47:419–448, 1978
Dignam JD, Lebovitz RM, Roeder RG: Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11:1475–1489, 1983
Hennighausen L, Lubon H: Interaction of protein with DNAin vitro. Meth Enzymol 152:721–735, 1987
Maniatis T, Fritsch EF, and Sambrook J: A laboratory manual. Cold Spring Harbor University Press, Cold Spring Harbor, New York, 1982
Cooney M, Czermuszewicz G, Postel E, Flint S, Hogan M: Site-specific oligonucleotide binding represses transcription of the humanc-myc genein vitro. Science 241:456–459, 1988
Lelong J-C, Prevost G, Lee K, Crepin M:In vitro characterization of tissue-specific nuclear proteins preferentially bound to the mouse β-globin gene during MEL cell terminal differentiation. Biochemistry 28:4594–4600, 1989
Jackson PD, Evans T, Nickol JM, Felsenfeld G: Developmental modulation of protein binding to β-globin gene regulatory sites within chicken erythrocyte nuclei. Genes Dev 3:1860–1873, 1989
Andrews NC, Erdjument-Bromage H, Davidson MB, Tempst P, Orkin SH: Erythroid transcriptional factor NF-E2 is a haemotopoietic-specific basic-leucine zipper protein. Nature 362:722–728, 1993
Harrington RE, Winicov I: New concepts in protein-DNA recognition: sequence directed DNA bending and flexibility. In: W.E. Cohn and K. Moldave (eds). Progress in Nucleic Acid Research and Molecular Biology. Academic Press, 1994, pp 195–270
Roberts RW, Crothers DM: Stability and properties of double and triple helices: dramatic effects of RNA or DNA backbone composition. Science 258:1463–1466, 1992
Postel EH, Flint SJ, Kessler DJ, Hogan ME: Evidence that a triplexforming oligodeoxyribonucleotide binds to thec-myc promoter in HeLa cells reducingc-myc mRNA levels. Proc Natl Acad Sci USA 88:8227–8231, 1991
Plumb M, Frampton J, Wainwright H, Walker M, Macleod K, Goodwin G, Harrison P: GAGAAG; acis-control region binding an erythroid-specific nuclear factor with a role in globin and non-globin gene expression. Nucleic Acids Res 17:73–92, 1989
Reddy PMS, Shen C-KJ: Erythroid differentiation of mouse erythroleukemia cells results in reorganization of protein-DNA complexes in the mouse βmaj globin promoter but not its distal enhancer. Mol Cell Biol 13:1093–1103, 1993
Sheffery M, Kim CG, Barnhart KM: Purification of four erythroid cell proteins that bind the promoters of the murine globin genes. In: G. Stamatoyannopoulos and A. W. Nienhuis (eds). Hemoglobin Switching Part A: Transcriptional Regulation. A.R. Liss Inc, New York, 1989, pp 343–357
Balcarek JM, McMorris AF: DNase I hypersensitive sites of globin genes of uninduced Friend enrythroleukemia cells and changes during induction with dimethylsulfoxide. J Biol Chem 258:10622–10628, 1983
Emerson BM, Nickol J, Fong TC: Erythroid-specific activation and derepression of the chick β-globin promoterin vitro. Cell 57: 1189–1200, 1989
Berg PE, Williams DM, Qian R-L, Cohen RB, Cao S-X, Mittelman M, Schechter AN: A common protein binds to two silencers 5′ to the human β-globin gene. Nucl Acids Res 17:8833–8852, 1989
Rahuel C, Vinit M-A, Lemarchandel V, Cartron J-P, Romeo P-H: Erythroid-specific activity of the gycophorin B promoter requires GATA-1 mediated displacement of a repressor. EMBO J 11: 4095–4102, 1992
Bentley DL, Groudine M: A block to elongation is largely responsible for decreased transcription ofc-myc in differentiated HL 60 cells. Nature 321:702–706, 1986
Theunissen O, Rudt F, Guddat U, Mentzel H, Pieler T: RNA and DNA binding zinc fingers in Xenopus TFIIIA. Cell 71:679–690, 1992
Clemens KR, Liao X, Wolf V, Wright PE, Gottesfeld JM: Definition of the binding sites of individual zinc fingers in the transcription factor IIIA-5S RNA gene complex. Proc Natl Acad Sci USA 89: 10822–10826, 1992
Schneider CA, Lim RW, Terwilliger E, Herschman HR: Epidermal growth factor-nonresponsive 3T3 variants do not contain epidermal growth factor receptor-related antigens or mRNA. Proc Natl Acad Sci USA 83:333–336, 1986
Cleghon VG, Klessig DF: Association of the adenovirus DNA-binding protein with RNA bothin vitro andin vivo. Proc Natl Acad Sci USA 83:8947–8951, 1986
Berkhout B, Gatignol A, Rabson AB, Jeang K-T: TAR-Independent activation of the HIV-1 LTR: evidence thatTat requires specific regions of the promoter. Cell 62:757–767, 1990
Dingwall C, Ernberg I, Gait MJ, Green MS, Heaphy S, Karn J, Lowe AD, Singh M, Skinner M, Valerio R: Human immunodeficiency virus 1 tat protein bindstrans-activation-responsive region (TAR) RNAin vitro. Proc Natl Acad Sci USA 86:6925–6929, 1989
Rastinejad F, Blau HM: Genetic complementation reveals a novel regulatory role for 3′ untranslated regions in growth and differentiation. Cell 72:903–917, 1993
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Sun-Hoffman, L., Winicov, I. Nuclear proteins that interact with the βmaj globin promoter start to accumulate in MEL cells within 12 hours of induction and RNA copies of the promoter successfully compete their bindingin vitro . Mol Cell Biochem 145, 159–168 (1995). https://doi.org/10.1007/BF00935488
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DOI: https://doi.org/10.1007/BF00935488